Poly(phosphoesters) for delivery of nucleic acids

ABSTRACT

Disclosed are polymers comprising the moiety A, which is a moiety of formula I: and pharmaceutically acceptable salts thereof, wherein R, R 1 , R 2 , L, n1 and n2 are as defined herein. These polymers are useful for delivering nucleic acids to subject. These polymers and pharmaceutically acceptable compositions comprising such polymers and nucleic acids can be useful for treating various diseases, disorders and conditions.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a 35 U.S.C. § 371 National Stage Application of International Application No. PCT/US18/36920, filed Jun. 11, 2018, which claims benefit of U.S. Provisional Application No. 62/518,313, filed Jun. 12, 2017, which is hereby incorporated by reference in its entirety.

SEQUENCE LISTING

The present specification makes reference to a Sequence Listing provided in electronic form as an ASCII.txt file named “MRT-1248WO_ST25.txt” that was generated on Jun. 11, 2018, and is 3,013 bytes in size.

BACKGROUND

Nucleic acids have been explored as a potential therapeutic option for certain disease states. In particular, ribonucleic acid (RNA) interference (RNAi) using, for example, small interfering RNA (siRNA), short hairpin RNA (shRNA), and antisense RNA (aRNA) approaches have been the subject of significant research and clinical development, for example, for binding to messenger RNA (mRNA), long non-coding RNA (lncRNA), micro-RNA (miRNA) and other endogenous targets. More recently, administration of mRNA, multimeric coding nucleic acid (MCNA), polymeric coding nucleic acid (PCNA), guide RNA (gRNA) and CRISPR RNA (crRNA) have been investigated as possible treatments of various diseases. However, the delivery of nucleic acids as therapeutics remains a challenge.

SUMMARY

The present invention provides, among other things, polymers useful in delivering nucleic acids as a therapeutic. The invention is based, in part, on the surprising discovery that the polymers described herein provide safe and efficient delivery of nucleic acids, such as mRNA. In particular, the polymers of the present invention may provide one or more advantages over other polymers. For example, as described in more detail herein, the polymers of the present invention may provide enhanced endosomal release of the nucleic acid being delivered by the polymer. Additionally, the biodegradable nature of the polymer may provide greater patient tolerability than other polymeric delivery vehicles. Thus, the polymers of the present invention may provide more potent and/or safer nucleic acid delivery for the treatment of a variety of diseases.

In some aspects, the present invention provides a polymer comprising a repeating unit that is:

or a pharmaceutically acceptable salt thereof, wherein:

-   -   each instance of R¹ independently is —H or a C₁-C₂₀ aliphatic         group;     -   each instance of R² independently is —H or a C₁-C₂₀ aliphatic         group;     -   each instance of R³ independently is —H or a C₁-C₂₀ aliphatic         group;     -   X is independently S or O;     -   n1 is an integer from 1 to 6;     -   n2 is an integer from 1 to 6;     -   R is N or CH; and     -   L is C₁₋₆ alkyl or a protonatable group provided that when R is         CH then L is a protonatable group, wherein the protonatable         group comprises at least one 1° amino, 2° amino, 3° amino or         imidazolyl group.

In some aspects, the present invention provides methods of preparing the polymers of the present invention.

In some aspects, the present invention provides a pharmaceutical composition (a “provided composition”) comprising a polymer of the present invention and one or more nucleic acids or polynucleotides.

In some aspects, the present invention provides methods of treating a disease in a subject comprising administering to the subject a provided composition.

Definitions

In order for the present invention to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. The publications and other reference materials referenced herein to describe the background of the invention and to provide additional detail regarding its practice are hereby incorporated by reference for all purposes.

As used in this Specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.

Unless specifically stated or obvious from context, as used herein, the term “or” is understood to be inclusive and covers both “or” and “and”.

The terms “e.g.,” and “i.e.” as used herein, are used merely by way of example, without limitation intended, and should not be construed as referring only those items explicitly enumerated in the specification.

The terms “or more”, “at least”, “more than”, and the like, e.g., “at least one” are understood to include but not be limited to at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 or more than the stated value. Also included is any greater number or fraction in between.

The terms “plurality”, “at least two”, “two or more”, “at least second”, and the like, are understood to include but not limited to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 or more. Also included is any greater number or fraction in between.

Throughout the specification the word “comprising,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

Approximately or about: As used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

Amino acid: As used herein, the term “amino acid,” in its broadest sense, refers to any compound and/or substance that can be incorporated into a polypeptide chain. In some embodiments, an amino acid has the general structure H₂N—C(H)(R)—COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a modified amino acid; in some embodiments, an amino acid is a d-amino acid; in some embodiments, an amino acid is an I-amino acid. “Standard amino acid” refers to any of the twenty standard I-amino acids commonly found in naturally occurring peptides. “Nonstandard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. As used herein, “modified amino acid” encompasses a chemically modified amino acids, including but not limited to salts, amino acid derivatives (such as amides), and/or substitutions. Amino acids, including carboxy- and/or amino-terminal amino acids in peptides, can be modified by methylation, amidation, acetylation, protecting groups, and/or substitution with other chemical groups that can change the peptide's circulating half-life without adversely affecting their activity. Amino acids may participate in a disulfide bond. Amino acids may comprise one or posttranslational modifications, such as association with one or more chemical entities (e.g., methyl groups, acetate groups, acetyl groups, phosphate groups, formyl moieties, isoprenoid groups, sulfate groups, polyethylene glycol moieties, lipid moieties, carbohydrate moieties, biotin moieties, etc.). The term “amino acid” is used interchangeably with “amino acid residue,” and may refer to a free amino acid and/or to an amino acid residue of a peptide. It will be apparent from the context in which the term is used whether it refers to a free amino acid or a residue of a peptide.

Animal: As used herein, the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to humans, at any stage of development. In some embodiments, “animal” refers to non-human animals, at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, and/or a pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and/or worms. In some embodiments, an animal may be a transgenic animal, genetically-engineered animal, and/or a clone.

Biologically active: As used herein, the term “biologically active” refers to a characteristic of any agent that has activity in a biological system, and particularly in an organism. For instance, an agent that, when administered to an organism, has a biological effect on that organism, is considered to be biologically active.

Delivery: As used herein, the term “delivery” refers to delivery of a composition to an organism, including but not limited to an animal or human. For delivery to a multi-tissue organism, delivery encompasses both local and systemic delivery. For example, delivery of a nucleic acid such as mRNA encompasses situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and retained within the target tissue (also referred to as “local distribution” or “local delivery”), and situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and secreted into patient's circulation system (e.g., serum) and systematically distributed and taken up by other tissues (also referred to as “systemic distribution” or “systemic delivery).

Expression: As used herein, “expression” of a nucleic acid sequence refers to one or more of the following events: (1) production of an mRNA template from a DNA sequence (e.g., by transcription); (2) processing of an mRNA transcript (e.g., by splicing, editing, 5′ cap formation, and/or 3′ end formation); (3) translation of an mRNA into a polypeptide or protein; and/or (4) post-translational modification of a polypeptide or protein. In this application, the terms “expression” and “production,” and grammatical equivalent, are used inter-changeably.

Functional: As used herein, a “functional” biological molecule is a biological molecule in a form in which it exhibits a property and/or activity by which it is characterized.

Half-life: As used herein, the term “half-life” is the time required for a quantity such as nucleic acid or protein concentration or activity to fall to half of its value as measured at the beginning of a time period.

Improve, increase, or reduce: As used herein, the terms “improve,” “increase” or “reduce,” or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control sample or subject (or multiple control samples or subjects) in the absence of the treatment described herein. A “control subject” is a subject afflicted with the same form of disease as the subject being treated, who is about the same age as the subject being treated.

In Vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc., rather than within a multi-cellular organism.

In Vivo: As used herein, the term “in vivo” refers to events that occur within a multi-cellular organism, such as a human and a non-human animal. In the context of cell-based systems, the term may be used to refer to events that occur within a living cell (as opposed to, for example, in vitro systems).

Isolated: As used herein, the term “isolated” refers to a substance and/or entity that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature and/or in an experimental setting), and/or (2) produced, prepared, and/or manufactured by the hand of man. Isolated substances and/or entities may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which they were initially associated. In some embodiments, isolated agents are about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure. As used herein, a substance is “pure” if it is substantially free of other components. As used herein, calculation of percent purity of isolated substances and/or entities should not include excipients (e.g., buffer, solvent, water, etc.).

messenger RNA (mRNA): As used herein, the term “messenger RNA (mRNA)” or “mRNA” refers to a polynucleotide that encodes at least one polypeptide. mRNA as used herein encompasses both modified and unmodified RNA. mRNA may contain one or more coding and non-coding regions. mRNA can be purified from natural sources, produced using recombinant expression systems or plasmid-based expression systems or in vitro transcription (IVT) systems, and optionally purified and/or modified, or produced by chemical synthesis. In some embodiments, for example, for mRNA produced using recombinant or plasmid-based expression systems or produced by chemical synthesis, the mRNA coding region, the mRNA non-coding region, or both the mRNA coding and non-coding regions, can include a unique nucleic acid sequence (having modified or unmodified nucleic acids), i.e., that is different from natural and/or known nucleic acid sequences for that mRNA. For example, in some embodiments, the mRNA coding region can include one or more codons that have a different triplet nucleic acid sequence than the triplet nucleic acid sequence of the corresponding codons in the corresponding natural or known mRNA (referred to herein as a “codon-optimized” sequence). An mRNA sequence is presented in the 5′ to 3′ direction unless otherwise indicated. In some embodiments, “mRNA” can encompass both unmodified RNA and modified mRNA (mmRNA). The modifications in mmRNA can comprise nucleoside analogs such as analogs having one or more chemically modified bases or sugars, backbone modifications, or other modifications described herein. In some embodiments, an mRNA is or comprises natural nucleosides (e.g., adenosine, guanosine, cytidine, uridine); nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); intercalated bases; modified sugars (e.g., 2′-fluororibose, ribose, 2′-deoxyribose, arabinose, and hexose); and/or modified phosphate groups (e.g., phosphorothioates and 5′-N-phosphoramidite linkages).

Nucleic acid: As used herein, the term “nucleic acid,” in its broadest sense, refers to any compound and/or substance that is or can be incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and/or substance that is or can be incorporated into a polynucleotide chain via a phosphodiester linkage. In some embodiments, “nucleic acid” refers to individual nucleic acid residues (e.g., nucleotides and/or nucleosides). In some embodiments, “nucleic acid” refers to a polynucleotide chain comprising individual nucleic acid residues. In some embodiments, “nucleic acid” encompasses ribonucleic acids (RNA), including but not limited to any one or more of interference RNAs (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA (mmRNA), long non-coding RNA (lncRNA), micro-RNA (miRNA) multimeric coding nucleic acid (MCNA), polymeric coding nucleic acid (PCNA), guide RNA (gRNA) and CRISPR RNA (crRNA). In some embodiments, “nucleic acid” encompasses deoxyribonucleic acid (DNA), including but not limited to any one or more of single-stranded DNA (ssDNA), double-stranded DNA (dsDNA) and complementary DNA (cDNA). In some embodiments, “nucleic acid” encompasses both RNA and DNA. In embodiments, DNA may be in the form of antisense DNA, plasmid DNA, parts of a plasmid DNA, pre-condensed DNA, a product of a polymerase chain reaction (PCR), vectors (e.g., P1, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives of these groups. In embodiments, RNA may be in the form of messenger RNA (mRNA), ribosomal RNA (rRNA), signal recognition particle RNA (7 SL RNA or SRP RNA), transfer RNA (tRNA), transfer-messenger RNA (tmRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), SmY RNA, small Cajal body-specific RNA (scaRNA), guide RNA (gRNA), ribonuclease P (RNase P), Y RNA, telomerase RNA component (TERC), spliced leader RNA (SL RNA), antisense RNA (aRNA or asRNA), cis-natural antisense transcript (cis-NAT), CRISPR RNA (crRNA), long noncoding RNA (lncRNA), micro-RNA (miRNA), piwi-interacting RNA (piRNA), small interfering RNA (siRNA), transacting siRNA (tasiRNA), repeat associated siRNA (rasiRNA), 73K RNA, retrotransposons, a viral genome, a viroid, satellite RNA, or derivatives of these groups. In some embodiments, a nucleic acid is a mRNA encoding a protein such as an enzyme.

Patient: As used herein, the term “patient” or “subject” refers to any organism to which a provided composition may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, a patient is a human. A human includes pre- and post-natal forms.

Pharmaceutically acceptable: The term “pharmaceutically acceptable”, as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

Pharmaceutically acceptable salt: Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N⁺(C₁₋₄ alkyl)₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium. quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, sulfonate and aryl sulfonate. Further pharmaceutically acceptable salts include salts formed from the quarternization of an amine using an appropriate electrophile, e.g., an alkyl halide, to form a quarternized alkylated amino salt.

Systemic distribution or delivery: As used herein, the terms “systemic distribution,” “systemic delivery,” or grammatical equivalent, refer to a delivery or distribution mechanism or approach that affect disparate compartments or tissues of the entire body or of an entire organism. Typically, systemic distribution or delivery is accomplished via body's circulation system, e.g., blood stream. Compared to the definition of “local distribution or delivery.”

Subject: As used herein, the term “subject” refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). A human includes pre- and post-natal forms. In many embodiments, a subject is a human being. A subject can be a patient, which refers to a human presenting to a medical provider for diagnosis or treatment of a disease. The term “subject” is used herein interchangeably with “individual” or “patient.” A subject can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder.

Substantially: As used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

Target tissues: As used herein, the term “target tissues” refers to any tissue that is affected by a disease to be treated. In some embodiments, target tissues include those tissues that display disease-associated pathology, symptom, or feature.

Therapeutically effective amount: As used herein, the term “therapeutically effective amount” of a therapeutic agent means an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and/or condition, to treat, diagnose, prevent, and/or delay the onset of the symptom(s) of the disease, disorder, and/or condition. It will be appreciated by those of ordinary skill in the art that a therapeutically effective amount is typically administered via a dosing regimen comprising at least one unit dose.

Treating: As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of and/or reduce incidence of one or more symptoms or features of a particular disease, disorder, and/or condition. Treatment may be administered to a subject who does not exhibit signs of a disease and/or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease.

Aliphatic: As used herein, the term aliphatic refers to both C₁-C₄₀ hydrocarbons and includes both saturated and unsaturated hydrocarbons. An aliphatic may be linear, branched, or cyclic. For example, C₁-C₂₀ aliphatics can include C₁-C₂₀ saturated alkyls (e.g., linear or branched C₁-C₂₀ saturated alkyls), C₂-C₂₀ alkenyls (e.g., linear or branched C₄-C₂₀ dienyls, linear or branched C₆-C₂₀ trienyls, and the like), and C₂-C₂₀ alkynyls (e.g., linear or branched C₂-C₂₀ alkynyls). aliphatics can include C₃-C₂₀ cyclic aliphatics (e.g., C₃-C₂₀ cycloalkyls, C₄-C₂₀ cycloalkenyls, or C₈-C₂₀ cycloalkynyls). The term “alkylene,” as used herein, represents a saturated divalent straight or branched chain hydrocarbon group and is exemplified by methylene, ethylene, isopropylene and the like. In certain embodiments, the aliphatic may comprise one or more cyclic aliphatic and/or one or more heteroatoms such as oxygen, nitrogen, or sulfur and may optionally be substituted with one or more substituents such as alkyl, halo, alkoxyl, hydroxy, amino, aryl, ether, ester or amide. For example, an aliphatic may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, —CO₂R′, —CN, —OH, —OR′, —NH₂, —NHR′, —N(R′)₂, —SR′ or —SO₂R′, wherein each instance of R′ independently is C₁₋₃ alkyl. In certain embodiments, the aliphatic is unsubstituted. In certain embodiments, the aliphatic does not include any heteroatoms.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Polymers—Moiety A

In some aspects, the present invention provides a polymer comprising a repeating unit that is:

or a pharmaceutically acceptable salt thereof, wherein:

-   -   each instance of R¹ independently is —H or a C₁-C₂₀ aliphatic         group;     -   each instance of R² independently is —H or a C₁-C₂₀ aliphatic         group;     -   each instance of R³ independently is —H or a C₁-C₂₀ aliphatic         group;     -   X is independently S or O;     -   n1 is an integer from 1 to 6;     -   n2 is an integer from 1 to 6;     -   R is N or CH; and     -   L is C₁₋₆ alkyl or a protonatable group provided that when R is         CH then L is a protonatable group, wherein the protonatable         group comprises at least one 1° amino, 2° amino, 3° amino or         imidazolyl group.

In some embodiments, R¹ independently is —H, —CH₃, or —CH₂CH₃.

In some embodiments, R² independently is —H, —CH₃, or —CH₂CH₃.

In some embodiments, R³ independently is —H, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, CH(CH₃)₂, or

C(CH₃)₃.

In some embodiments, each instance of R¹ and R² independently is —H or —CH₃. In some embodiments, each instance of R¹ and R² is —H, —CH₃ or —CH₂CH₃. In some embodiments, each instance of R¹ and R² is —CH₃. In some embodiments, each instance of R¹ and R² is —H.

In some embodiments, X is independently O.

In some embodiments, X is independently S.

In some embodiments, n1 is 1, 2 or 3; or n1 is 2, 3 or 4; or n1 is 3, 4 or 5; or n1 is 4, 5 or 6; or n1 is 1 or 2; or n1 is 2 or 3; or n1 is 3 or 4; or n1 is 4 or 5; or n1 is 5 or 6; or n1 is 1; or n1 is 2; or n1 is 3; or n1 is 4; or n1 is 5; or n1 is 6.

In some embodiments, n2 is 1, 2 or 3; or n2 is 2, 3 or 4; or n2 is 3, 4 or 5; or n2 is 4, 5 or 6; or n2 is 1 or 2; or n2 is 2 or 3; or n2 is 3 or 4; or n2 is 4 or 5; or n2 is 5 or 6; or n2 is 1; or n2 is 2; or n2 is 3; or n2 is 4; or n2 is 5; or n2 is 6.

In some embodiments,

-   n1 is 1, 2 or 3; or n1 is 2, 3 or 4; or n1 is 3, 4 or 5; or n1 is 4,     5 or 6; or n1 is 1 or 2; or n1 is 2 or 3; or n1 is 3 or 4; or n1 is     4 or 5; or n1 is 5 or 6; or n1 is 1; or n1 is 2; or n1 is 3; or n1     is 4; or n1 is 5; or n1 is 6; -   n2 is 1, 2 or 3; or n2 is 2, 3 or 4; or n2 is 3, 4 or 5; or n2 is 4,     5 or 6; or n2 is 1 or 2; or n2 is 2 or 3; or n2 is 3 or 4; or n2 is     4 or 5; or n2 is 5 or 6; or n2 is 1; or n2 is 2; or n2 is 3; or n2     is 4; or n2 is 5; or n2 is 6; and -   each instance of R¹ and R² independently is —H or —CH₃; or each     instance of R¹ and R² is —H, —CH₃ or —CH₂CH₃; or each instance of R¹     and R² is —CH₃; or each instance of R¹ and R² is —H.

In some embodiments, R is CH.

In some embodiments, R is N.

In some embodiments, R is N and L is C₁₋₆ alkyl.

In some embodiments, L is a protonatable group. A suitable protonatable group does not significantly interfere with the delivery of nucleic acid using the polymer of the present invention. Generally, a suitable protonatable group is a C₁₋₁₀ hydrocarbon additionally containing at least one 1° amino, 2° amino, 3° amino or imidazolyl group. In some embodiments, non-protonatable linkages such as esters and ethers may also be present. In some embodiments, sidechains of the naturally occurring amino acids may also be present.

In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 4.5 to about 11.5. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 5 to about 11. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 6 to about 10. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 7 to about 9. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 8. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of 10.

In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 5 to about 7. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 6 or 7. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 8 to about 11. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 9 or 10. In some embodiments, L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 8.

In some embodiments, R is CH and L is a protonatable group. In some embodiments, R is N and L is a protonatable group.

In some embodiments, a repeating unit is

-   or a pharmaceutically acceptable salt thereof, wherein: -   L is C₁₋₆ alkyl; and -   each of R, R¹, R², R³, X, n1 and n2 is as defined above, both singly     and in combination.

In some embodiments, L is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-Bu, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, L is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, L is —CH₃, Et or Pr. In some embodiments, L is —CH₃. In some embodiments, L is Et. In some embodiments, L is Pr. In some embodiments, L is ^(i)Pr. In some embodiments, L is Bu.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, X, L, n1 and n2 is as defined and     described in embodiments above, both singly and in combination.

In some embodiments, the repeating unit is:

-   wherein each of R¹, R², R³, X, L, n1 and n2 is as defined and     described in embodiments above, both singly and in combination.

In some embodiments, a repeating unit is:

-   or a pharmaceutically acceptable salt thereof, wherein: -   each of R⁴ and R⁵ independently is —H or a C₁-C₂₀ aliphatic group     (e.g., a C1-6 alkyl); and -   each of R, R¹, R², R³, X, n1 and n2 is as defined above, both singly     and in combination.

In some embodiments, R⁴ independently is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-Bu, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R⁴ is —H, —CH₃, Et, Pr, ^(i)Pr or Bu; or R⁴ is —CH₃, Et, Pr, ^(i)Pr or Bu; or R⁴ is —H, —CH₃, Et or Pr; or R⁴ is —CH₃, Et or Pr; or R⁴ is —CH₃; or R⁴ is Et; or R⁴ is Pr.

In some embodiments, R⁵ independently is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-Bu, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R⁵ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-Bu, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R⁵ is —CH₃, Et, Pr, ^(i)Pr or Bu; or R⁴ is —CH₃, Et or Pr; or R⁵ is —CH₃; or R⁵ is Et; or R⁵ is Pr.

In some embodiments, R⁴ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-Bu, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R⁴ is —H, —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R⁴ is —CH₃, Et, Pr, iPr or Bu. In some embodiments, R⁴ is —H, —CH₃, Et or Pr. In some embodiments, R⁴ is —CH₃, Et or Pr. In some embodiments, R⁴ is —CH₃. In some embodiments, R⁴ is Et. In some embodiments, R⁴ is Pr.

In some embodiments, R⁵ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-Bu, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R⁵ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R⁵ is —CH₃, Et or Pr. In some embodiments, R⁵ is —CH₃. In some embodiments, R⁵ is Et. In some embodiments, R⁵ is Pr.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁴, R⁵, X, n1 and n2 is as defined and     described in embodiments above.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁴, R⁵, X, n1 and n2 is as defined and     described in embodiments above, both singly and in combination.

In some embodiments, a repeating unit is:

-   or a pharmaceutically acceptable salt thereof, wherein: -   n3 is 1, 2, 3 or 4; -   R⁷ is halo, C₁₋₄ alkyl, C₁₋₄ haloalkyl, —OH, —OR^(J1), —NH₂,     —NHR^(J1), —N(R^(J1))₂, —C(O)R^(J1), —CN or —NO₂, wherein each     instance of R^(J1) is independently C₁₋₃ alkyl; -   R⁶ is —H or a C₁-C₂₀ aliphatic group (e.g., a C₁₋₆ alkyl); and -   each of R, R¹, R², R³, X, n1, and n2 is as defined above, both     singly and in combination.

In some embodiments, R⁶ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl, or hexyl; or R⁶ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl, or hexyl; or R⁶ is —CH₃, Et, Pr, ^(i)Pr, or Bu; or R⁶ is —CH₃, Et, or Pr; or R⁶ is —CH₃.

In some embodiments, R⁷ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂; or R⁷ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂; or R⁷ is —CH₃, Et or Pr; or R⁷ is Pr.

In some embodiments, n3 is 1, 2 or 3; or n3 is 2, 3 or 4; or n3 is 2 or 3; or n3 is 2; or n3 is 3.

In some embodiments, R⁶ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl, or hexyl. In some embodiments, R⁶ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl, or hexyl. In some embodiments, R⁶ is —CH₃, Et, Pr, ^(i)Pr, or Bu. In some embodiments, R⁶ is —CH₃, Et, or Pr. In some embodiments, R⁶ is —CH₃.

In some embodiments, R⁷ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂. In some embodiments, R⁷ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂. In some embodiments, R⁷ is —CH₃, Et or Pr. In some embodiments, R⁷ is Pr.

In some embodiments, n3 is 1, 2 or 3. In some embodiments, n3 is 2, 3 or 4. In some embodiments, n3 is 2 or 3. In some embodiments, n3 is 2. In some embodiments, n3 is 3.

In some embodiments, a repeating unit is

-   wherein each of R¹, R², R³, R⁶, R⁷, X, n1, n2 and n3 is as defined     and described in embodiments above, both singly and in combination.

In some embodiments, a repeating unit is

-   wherein each of R¹, R², R³, R⁶, R⁷, X, n1, n2, and n3 is as defined     and described in embodiments above, both singly and in combination.

In some embodiments, a repeating unit is:

-   or a pharmaceutically acceptable salt thereof, wherein: -   each of R⁸, R⁹, R¹⁰ and R¹¹ independently is a C₁-C₂₀ aliphatic     (e.g., a C₁₋₆ aliphatic, C₁₋₄ haloalkyl or C₃₋₆ cycloalkyl, each of     which is optionally substituted with 1-4 occurrences of R*); -   each instance of R* independently is halogen, —CO₂R^(J1), —CN, —OH,     —OR^(J1), —NH₂, —NHR^(J1), —N(R^(J1))₂, —SR^(J1) or —SO₂R^(J1),     wherein each instance of R^(J1) independently is C₁₋₃ alkyl; and -   each of R¹, R², R³, X, n1 and n2 is as defined above for formula     (I), both singly and in combination.

In some embodiments, R⁸ is —H, —CH₃, Et, Pr, Bu, —(CH₂)₃NH₂, —(CH₂)₄NH₂ or —(CH₂)₅NH₂; or R⁸ is —H, —CH₃, Et, —(CH₂)₄NH₂, or —(CH₂)₅NH₂; or R⁸ is —H or —(CH₂)₄NH₂; or R⁸ is —(CH₂)₄NH₂.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂; or R⁹ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂; or R⁹ is —CH₃, Et or Pr; or R⁹ is Pr.

In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R¹⁰ is —CH₃, Et, Pr, ^(i)Pr or Bu; or R¹⁰ is —CH₃, Et or Pr; or R¹⁰ is —CH₃.

In some embodiments, R¹¹ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; or R¹¹ is H, —CH₃, Et, Pr or cyclopropyl; or R¹¹ is H, —CH₃ or Et; or R¹¹ is —H.

In some embodiments, R⁸ is —H, —CH₃, Et, Pr, Bu, —(CH₂)₃NH₂, —(CH₂)₄NH₂ or —(CH₂)₅NH₂. In some embodiments, R⁸ is —H, —CH₃, Et, —(CH₂)₄NH₂, or —(CH₂)₅NH₂. In some embodiments, R⁸ is —H or —(CH₂)₄NH₂. In some embodiments, R⁸ is —(CH₂)₄NH₂.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂. In some embodiments, R⁹ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂. In some embodiments, R⁹ is —CH₃, Et or Pr. In some embodiments, R⁹ is Pr.

In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R¹⁰ is —CH₃, Et or Pr. In some embodiments, R¹⁰ is —CH₃.

In some embodiments, R¹¹ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, R¹¹ is H, —CH₃, Et, Pr or cyclopropyl. In some embodiments, R¹¹ is H, —CH₃ or Et. In some embodiments, R¹¹ is —H.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁸, R⁹, R¹⁰, R¹¹, X, n1 and n2 is as     defined and described in embodiments above, both singly and in     combination.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁸, R⁹, R¹⁰, R¹¹, X, n1 and n2 is as     defined and described in embodiments above, both singly and in     combination.

In some embodiments, a repeating unit is:

-   or a pharmaceutically acceptable salt thereof, wherein: -   B is a 5-membered heterocycle having 1 or 2 ring nitrogen atoms; -   n4 is 1, 2, 3 or 4; -   each instance of R¹² independently is selected from halo, C₁₋₄     alkyl, C₁₋₄ haloalkyl, —OH, —OR^(J1), —NH₂,     -   —NHR^(J1), —N(R^(J1))₂, —C(O)R^(J1), —CN and —NO₂;     -   wherein each instance of R^(J1) is independently C₁₋₃ alkyl; -   n5 is 0, 1, 2 or 3; and -   each of R, R¹, R², R³, X, n1 and n2 is as defined above, both singly     and in combination.

In some embodiments, B is an aromatic heterocyclic ring.

In some embodiments, B is a non-aromatic heterocyclic ring.

In some embodiments, B is pyrazolyl, imidazolyl, pyrrolyl, triazolyl, or tertazolyl; or B is pyrazolyl or imidazolyl; or B is imidazolyl.

In some embodiments, R¹² is —CH₃, Et, Pr, Bu, ^(i)Pr, —F, —Cl, —CF₃, —OMe, —OEt, —OPr, —NH₂, —SO₂NH₂, —CN or —NO₂; or R¹² is —CH₃, —F, —Cl, —CF₃, —OMe or —OEt; or R¹² is —F, —CH₃ or —Cl; or R¹² is —CH₃.

In some embodiments, n4 is 0, 1 or 2; or n4 is 0 or 1; or n4 is 0.

In some embodiments, n5 is 0, 1 or 2; or n5 is 0 or 1; or n5 is 0.

In some embodiments, B is pyrazolyl, imidazolyl, pyrrolyl, triazolyl, or tertazolyl. In some embodiments, B is pyrazolyl or imidazolyl. In some embodiments, B is imidazolyl.

In some embodiments, R¹² is —CH₃, Et, Pr, Bu, ^(i)Pr, —F, —Cl, —CF₃, —OMe, —OEt, —OPr, —NH₂, —SO₂NH₂, —CN or —NO₂. In some embodiments, R¹² is —CH₃, —F, —Cl, —CF₃, —OMe or —OEt. In some embodiments, R¹² is —F, —CH₃ or —Cl. In some embodiments, R¹² is —CH₃.

In some embodiments, n4 is 0, 1 or 2. In some embodiments, n4 is 0 or 1. In some embodiments, n4 is 0.

In some embodiments, n5 is 0, 1 or 2. In some embodiments, n5 is 0 or 1. In some embodiments, n5 is 0.

In some embodiments, n5 is 0, 1, or 2; and R¹² is —F or —CH₃. In some embodiments, n5 is 1 and R¹² is —CH₃.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R¹², B, X, n1, n2, n4 and n5 is as     defined and described in embodiments above, both singly and in     combination.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R¹², X, B, n1, n2, n4 and n5 is as     defined and described in embodiments above, both singly and in     combination.

In some embodiments, a repeating unit is:

-   or a pharmaceutically acceptable salt thereof, wherein: -   X′ is —O— or —NR¹³—, wherein     -   R¹³ is —H or a C₁-C₂₀ aliphatic (e.g., a C₁₋₆ aliphatic or C₃₋₆         cycloalkyl); -   n4 is 0, 1, 2 or 3; -   each of R⁸, R⁹ and R¹⁰ is —H or is a C₁-C₂₀ aliphatic (e.g., a C₁₋₆     aliphatic, C₁₋₄ haloalkyl or C₃₋₆ cycloalkyl, each of which is     optionally substituted with 1-4 occurrences of R*); -   each instance of R* independently is halogen, —CO₂R^(J1), —CN, —OH,     —OR^(J1), —NH₂, —NHR^(J1), —N(R^(J1))₂, —S^(J1) or —SO₂R^(J1),     -   wherein each instance of R^(J1) independently is C₁₋₃ alkyl; and -   each of R, R¹, R², R³, X, n1 and n2 is as defined above, both singly     and in combination.

In some embodiments, X′ is —NH— or —O—; or X′ is —NR¹³—; or X′ is —NH—; or X′ is —O—.

In some embodiments, n4 is 1, 2 or 3; or n4 is 1 or 2; or n4 is 2 or 3; or n4 is 1.

In some embodiments, R⁸ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂; or R⁸ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂; or R⁸ is —CH₃, Et or Pr; or R⁸ is Pr.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R⁹ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R⁹ is —CH₃, Et, Pr, ^(i)Pr or Bu; or R⁹ is —CH₃, Et or Pr; or R⁹ is —CH₃.

In some embodiments, R¹⁰ is —H, —CH₃, Et, Pr, Bu, —(CH₂)₃NH₂, —(CH₂)₄NH₂ or —(CH₂)₅NH₂; or R¹⁹ is —H, —CH₃, Et, —(CH₂)₄NH₂ or —(CH₂)₅NH₂; or R¹⁰ is —H or —(CH₂)₄NH₂; or R¹⁰ is —(CH₂)₄NH₂.

In some embodiments, R¹³ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; or R¹³ is —H, —CH₃, Et, cyclopropyl or cyclobutyl; or R¹³ is —H, —CH₃ or cyclopropyl; or R¹³ is —H.

In some embodiments, X′ is —NH— or —O—. In some embodiments, X′ is —NR¹³. In some embodiments, X′ is —NH—. In some embodiments, X′ is —O—.

In some embodiments, n4 is 1, 2 or 3. In some embodiments, n4 is 1 or 2. In some embodiments, n4 is 2 or 3. In some embodiments, n4 is 1.

In some embodiments, R⁸ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂. In some embodiments, R⁸ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂. In some embodiments, R⁸ is —CH₃, Et or Pr. In some embodiments, R⁸ is Pr.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R⁹ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R⁹ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R⁹ is —CH₃, Et or Pr. In some embodiments, R⁹ is —CH₃.

In some embodiments, R¹⁰ is —H, —CH₃, Et, Pr, Bu, —(CH₂)₃NH₂, —(CH₂)₄NH₂ or —(CH₂)₅NH₂. In some embodiments, R¹⁰ is —H, —CH₃, Et, —(CH₂)₄NH₂ or —(CH₂)₅NH₂. In some embodiments, R¹⁰ is —H or —(CH₂)₄NH₂. In some embodiments, R¹⁰ is —(CH₂)₄NH₂.

In some embodiments, R¹³ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, R¹³ is —H, —CH₃, Et, cyclopropyl or cyclobutyl. In some embodiments, R¹³ is —H, —CH₃ or cyclopropyl. In some embodiments, R¹³ is —H.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁹, R¹⁰, n1, n2, X, and X′ is as defined     and described in embodiments above and below, both singly and in     combination.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂. In some embodiments, R⁹ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂. In some embodiments, R⁹ is —CH₃, Et or Pr. In some embodiments, R⁹ is Pr.

In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R¹⁰ is —CH₃, Et, or Pr. In some embodiments, R¹⁰ is —CH₃.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁹, R¹⁰, n1, n2, X, and X′ is as defined     and described in embodiments above and below, both singly and in     combination.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂. In some embodiments, R⁹ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂. In some embodiments, R⁹ is —CH₃, Et or Pr. In some embodiments, R⁹ is Pr.

In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R¹⁰ is —CH₃, Et, or Pr. In some embodiments, R¹⁰ is —CH₃.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁸, R⁹, R¹⁰, n1, n2, X, and X′ is as     defined and described in embodiments above and below, both singly     and in combination.

In some embodiments, R⁸ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂. In some embodiments, R⁸ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂. In some embodiments, R⁸ is —CH₃, Et or Pr. In some embodiments, R⁸ is Pr.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R⁹ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R⁹ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R⁹ is —CH₃, Et or Pr. In some embodiments, R⁹ is —CH₃.

In some embodiments, R¹⁰ is —H, —CH₃, Et, Pr, Bu, —(CH₂)₃NH₂, —(CH₂)₄NH₂ or —(CH₂)₅NH₂. In some embodiments, R¹⁰ is —H, —CH₃, Et, —(CH₂)₄NH₂ or —(CH₂)₅NH₂. In some embodiments, R¹⁰ is —H or —(CH₂)₄NH₂. In some embodiments, R¹⁰ is —(CH₂)₄NH₂.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁹, R¹⁰, R¹³, X, n1, and n2 is as     defined and described in embodiments of formula (VII), above, and in     embodiments of formula (VIId), below, both singly and in     combination.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt or —NEt₂. In some embodiments, R⁹ is —H, —CH₃, Et, Pr, —NH₂, —NHMe or —NMe₂. In some embodiments, R⁹ is —CH₃, Et or Pr. In some embodiments, R⁹ is Pr.

In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R¹⁰ is —CH₃, Et, or Pr. In some embodiments, R¹⁰ is —CH₃.

In some embodiments, R¹³ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, R¹³ is —H, —CH₃, Et, cyclopropyl or cyclobutyl. In some embodiments, R¹³ is —H, —CH₃ or cyclopropyl. In some embodiments, R¹³ is —H.

In some embodiments, a repeating unit is:

-   or a pharmaceutically acceptable salt thereof, wherein: -   R¹⁴ is a side chain of an amino acid (e.g., a naturally-occurring     amino acid); -   n4 is 0, 1, 2 or 3; -   each of R⁹ and R¹⁰ is —H or is a C₁-C₂₀ aliphatic (e.g., a C₁₋₆     aliphatic, C₁₋₄ haloalkyl or C₃₋₆ cycloalkyl, each of which is     optionally substituted with 1-4 occurrences of R*). -   each instance of R* independently is halogen, —CO₂R^(J1), —CN, —OH,     —OR^(J1), —NH₂, —NHR^(J1), —N(R^(J1))₂, —SR^(J1) or —SO₂R^(J1),     -   wherein each instance of R^(J1) independently is C₁₋₃ alkyl; and -   each of R, R¹, R², R³, X, n1 and n2 is as defined above, both singly     and in combination.

In some embodiments, n4 is 1, 2 or 3; or n4 is 1 or 2; or n4 is 2 or 3; or n4 is 1.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt, or —NEt₂; or R⁹ is —H, —CH₃, Et, Pr, —NH₂, —NHMe, or —NMe₂; or R⁹ is —CH₃, Et, or Pr; or R⁹ is Pr.

In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R¹⁰ is —CH₃, Et, Pr, ^(i)Pr or Bu; or R¹⁰ is —CH₃, Et or Pr; or R¹⁰ is —CH₃.

In some embodiments, R¹⁴ is a side chain of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, lysine, arginine or histidine; or R¹⁴ is a side chain of tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, lysine, arginine or histidine; or R¹⁴ is a side chain of lysine or histidine.

In some embodiments, n4 is 1, 2 or 3. In some embodiments, n4 is 1 or 2. In some embodiments, n4 is 2 or 3. In some embodiments, n4 is 1.

In some embodiments, R⁹ is —H, —CH₃, Et, Pr, Bu, —NH₂, —NHMe, —NMe₂, —NHEt, or —NEt₂. In some embodiments, R⁹ is —H, —CH₃, Et, Pr, —NH₂, —NHMe, or —NMe₂. In some embodiments, R⁹ is —CH₃, Et, or Pr. In some embodiments, R⁹ is Pr.

In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R¹⁰ is —CH₃, Et, Pr, ^(i)Pr or Bu. In some embodiments, R¹⁰ is —CH₃, Et or Pr. In some embodiments, R¹⁰ is —CH₃.

In some embodiments, R¹⁴ is a side chain of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, lysine, arginine or histidine. In some embodiments, R¹⁴ is a side chain of tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, lysine, arginine or histidine. In some embodiments, R¹⁴ is a side chain of lysine or histidine.

In some embodiments, R¹⁴ is —H, —CH₃, ^(i)Pr, ^(i)Bu, sec-butyl, —CH₂OH, —CH₂SH,

In some embodiments, R¹⁴ is —CH₂SH, —CH₂OH,

In some embodiments, R¹⁴ is

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁹, R¹⁰, R¹⁴, X, n1, and n2 is as     defined and described in embodiments above, both singly and in     combination.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R⁹, R¹⁰, R¹⁴, X, n1, and n2 is as     defined and described in embodiments above, both singly and in     combination.

In some embodiments, a repeating unit is:

-   or a pharmaceutically acceptable salt thereof, wherein: -   X′ is —O— or —NR¹³—, wherein     -   R¹³ is —H, C₁₋₆ aliphatic or C₃₋₆ cycloalkyl; -   n4 is 0, 1, 2 or 3; -   n¹⁵ is —H or C₁₋₆alkyl; -   n6 is 1, 2, 3 or 4; and -   each of R, R¹, R², R³, X, n1 and n2 is as defined above, both singly     and in combination.

In some embodiments, X is —O— or —NH—; or X is —O—; or X is —NH—.

In some embodiments, n4 is 1, 2 or 3; or n4 is 1 or 2; or n4 is 2 or 3; or n4 is 1.

In some embodiments, n6 is 1, 2, 3 or 4; or n6 is 2 or 3; or n6 is 2.

In some embodiments, R¹⁵ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl; or R¹⁵ is —H, —CH₃, Et, Pr, ^(i)Pr, or Bu; or R¹⁵ is —H, —CH₃, or Et, or Pr; or R¹⁵ is —H.

In some embodiments, X is —O— or —NH—. In some embodiments, X is —O—. In some embodiments X is —NH—.

In some embodiments, n4 is 1, 2 or 3. In some embodiments, n4 is 1 or 2. In some embodiments, n4 is 2 or 3. In some embodiments, n4 is 1.

In some embodiments, n6 is 1, 2, 3 or 4. In some embodiments, n6 is 2 or 3. In some embodiments n6 is 2.

In some embodiments, R¹⁵ is —H, —CH₃, Et, Pr, ^(i)Pr, Bu, sec-butyl, ^(i)Bu, tert-Bu, pentyl, tert-pentyl, isopentyl, sec-pentyl, 3-pentyl or hexyl. In some embodiments, R¹⁵ is —H, —CH₃, Et, Pr, ^(i)Pr, or Bu. In some embodiments, R¹⁵ is —H, —CH₃, or Et, or Pr. In some embodiments, R¹⁵ is —H.

In some embodiments, n6 is 2 or 3 and R¹⁵ is —H, —CH₃, Et or Pr. In some embodiments, n6 is 2 and R¹⁵ is —H or —CH₃. In some embodiments, n6 is 2 and R¹⁵ is —H.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R¹⁵, X, n1, n2 and n6 is as defined and     described in embodiments of above, both singly and in combination.

In some embodiments, a repeating unit is:

-   wherein each of R¹, R², R³, R¹⁵, X, n1, n2 and n6 is as defined and     described in embodiments of formula (IX), above, both singly and in     combination.

Examples of polymers of the present invention include those comprising

a repeating group that is

wherein:

-   -   each instance of R¹ independently is —H or a C₁-C₂₀ aliphatic         group;     -   each instance of R² independently is —H or a C₁-C₂₀ aliphatic         group;     -   each instance of R³ independently is —H or a C₁-C₂₀ aliphatic         group;     -   X is independently S or O;     -   n1 is an integer from 1 to 6;     -   n2 is an integer from 1 to 6; and

is any of the following:

In some embodiments, R¹ independently is —H, —CH₃ or —CH₂CH₃.

In some embodiments, R² independently is —H, —CH₃ or —CH₂CH₃.

In some embodiments, R³ independently is —H, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, CH(CH₃)₂, or C(CH₃)₃.

In some embodiments, X is independently O.

In some embodiments, X is independently S.

Polymers

In some aspects, the present invention provides a polymer comprising the moiety A:

A

, which is a repeating unit as defined above (a “provided polymer”).

In some embodiments, a provided polymer is a homopolymer. In some embodiments, a provided homopolymer comprises the structure:

wherein m is an integer such that the weight average molecular weight (Mw) of the polymer is about 10 kDa to about 60 kDa.

In some embodiments, m is an integer such that M_(w) is about 10 kDa to about 60 kDa, about 10 kDa to about 50 kDa, about 10 kDa to about 40 kDa, about 10 kDa to about 30 kDa about 15 kDa to about 60 kDa, about 15 kDa to about 50 kDa, about 15 kDa to about 40 kDa, or about 15 kDa to about 30 kDa.

In some embodiments, the moiety:

constitutes at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% of the total M_(w) of the polymer.

In some embodiments, a provided polymer is a copolymer.

In some embodiments, a copolymer is a controlled di-block copolymer. In some embodiments, a copolymer is a controlled tri-block copolymer. In some embodiments, a copolymer is a mixed polymer. In some embodiments, a copolymer is an alternating copolymer. In some embodiments, a copolymer is a random copolymer. In some embodiments, a copolymer is a graft copolymer. In some embodiments, a copolymer is a periodic copolymer. In some embodiments, a copolymer is a statistical copolymer. In embodiments a copolymer is linear. In some embodiments, a copolymer is a linear copolymer. In some embodiments, a copolymer is a branched copolymer.

In some embodiments, a provided copolymer comprises two different segments:

-   m₁ instances of     A     and m₂ instances of     B     , -   wherein: -   A is a repeating unit as described above; -   B is a structurally distinct repeating unit from A (e.g., B is a     moiety of formula (I) as described above but different from A); and -   m₁ and m₂ are integers such that the weight average molecular weight     (M_(w)) of the polymer is about 10 kDa to about 60 kDa.

In some embodiments, B is a repeating unit as described above but different from A.

In some embodiments, B does not comprise any ionizable group. In some embodiments, B is neutral. In some embodiments, B is uncharged. In some embodiments, B is negatively charged. In some embodiments, B is positively charged.

In some embodiments, B is a targeting group (e.g., a targeting group as described herein).

In some embodiments, B is biodegradable. In embodiments, B is non-biodegradable.

In some embodiments, B is a moiety that is a hydrocarbon (e.g., a saturated hydrocarbon or an unsaturated hydrocarbon).

In some embodiments, B is a moiety that is a lactide (e.g., L-lactide or D,L-lactide), glycolide, ethylene glycol, caprolactone, propylene fumarate, or a 2-oxazoline.

In some embodiments, m₁ and m₂ are integers such that M_(w) is about 10 kDa to about 60 kDa, about 10 kDa to about 50 kDa, about 10 kDa to about 40 kDa, about 10 kDa to about 30 kDa about 15 kDa to about 60 kDa, about 15 kDa to about 50 kDa, about 15 kDa to about 40 kDa, or about 15 kDa to about 30 kDa.

In some embodiments, the

m₁ instances of

A

and m₂ instances of

B

together constitute at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% of the total M_(w) of the polymer.

In some embodiments, a provided copolymer is an alternating copolymer. In some embodiments, a provided alternating copolymer comprises the structure:

wherein m′ is an integer such that the weight average molecular weight (Mw) of the polymer is about 10 kDa to about 60 kDa.

In some embodiments, m′ is an integer such that M_(w) is about 10 kDa to about 60 kDa, about 10 kDa to about 50 kDa, about 10 kDa to about 40 kDa, about 10 kDa to about 30 kDa about 15 kDa to about 60 kDa, about 15 kDa to about 50 kDa, about 15 kDa to about 40 kDa, or about 15 kDa to about 30 kDa.

In some embodiments, the moiety:

constitutes at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% of the total M_(w) of the polymer.

In some embodiments, a provided copolymer is an block copolymer. In some embodiments, a provided block copolymer comprises the structure:

wherein m₁ and m₂ are integers such that the weight average molecular weight (M_(w)) of the polymer is about 10 kDa to about 60 kDa.

In some embodiments, m₁ and m₂ are integers such that M_(w) is about 10 kDa to about 60 kDa, about 10 kDa to about 50 kDa, about 10 kDa to about 40 kDa, about 10 kDa to about 30 kDa about 15 kDa to about 60 kDa, about 15 kDa to about 50 kDa, about 15 kDa to about 40 kDa, or about 15 kDa to about 30 kDa.

In some embodiments, the moiety:

constitutes at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% of the total M_(w) of the polymer.

In some embodiments, a provided polymer comprises the moiety:

A

T,

wherein:

A is a repeating unit as described above; and

T is a targeting group.

In embodiments, a provided polymer has hydrogen endgroups.

In embodiments, a provided polymer has oxygen-containing endgroups (e.g., hydroxyl or a C₁₋₆ alkoxyl).

In embodiments, a provided polymer has alkyl-containing endgroups (e.g., a C₁₋₆ alkyl).

In some embodiments, the targeting group of a polymer is a moiety that may interact with a biological target of interest via a biological binding event, i.e., between complementary pairs of biological molecules. For example, a targeting group may comprise an entity such as biotin that specifically binds to a complementary entity, such as avidin or streptavidin. Other examples of interactions that occur between pairs of biological molecules including proteins, nucleic acids, glycoproteins, carbohydrates, hormones, and the like. Specific examples include an antibody/peptide pair, an antibody/antigen pair, an antibody fragment/antigen pair, an antibody/antigen fragment pair, an antibody fragment/antigen fragment pair, an antibody/hapten pair, an enzyme/substrate pair, an enzyme/inhibitor pair, an enzyme/cofactor pair, a protein/substrate pair, a nucleic acid/nucleic acid pair, a protein/nucleic acid pair, a peptide/peptide pair, a protein/protein pair, a small molecule/protein pair, a glutathione/GST pair, an anti-GFP/GFP fusion protein pair, a Myc/Max pair, a maltose/maltose binding protein pair, a carbohydrate/protein pair, a carbohydrate derivative/protein pair, a metal binding tag/metal/chelate, a peptide tag/metal ion-metal chelate pair, a peptide/NT A pair, a lectin/carbohydrate pair, a receptor/hormone pair, a receptor/effector pair, a complementary nucleic acid/nucleic acid pair, a ligand/cell surface receptor pair, a virus/ligand pair, a Protein A/antibody pair, a Protein G/antibody pair, a Protein L/antibody pair, an Fe receptor/antibody pair, a biotin/avidin pair, a biotin/streptavidin pair, a drug/target pair, a zinc finger/nucleic acid pair, a small molecule/peptide pair, a small molecule/protein pair, a small molecule/target pair, a carbohydrate/protein pair such as maltose/MBP (maltose binding protein), a small molecule/target pair, or a metal ion/chelating agent pair.

In some embodiments, the inclusion of a, S-acyl-2-thioethyl (SATE) protecting group is employed. Such a protecting group is readily cleavable through esterase enzymatic activity resulting in a liberated phosphate functional group (Scheme 1). Upon cleavage of the SATE group, the generation of a negatively charged phosphate moiety is realized. The resulting negative charge can counter-balance the cationic charge of the amino monomer moiety resulting in a zwitterionic region. Upon release of some SATE protecting groups, the polymer is partially neutralized, rendering its ability to bind nucleic acids diminished. Thus, partial release of the nucleic acid payload will occur. Upon release of all SATE protecting groups, the polymer is completely neutralized, rendering its ability to bind nucleic acids removed. Thus, full release of the nucleic acid payload will occur. In total, such a polymer allows for facile complexation with nucleic acids, efficient release of payload and biodegradability of itself.

In some embodiments, a branched poly(phosphoester) can be produced via reaction with any diol monomer with a reactive phosphate moiety without inclusion of the S-acyl-thioethanol group (Scheme 2). Further, a mixed branched poly(phosphoester) polymer can be synthesized incorporating both the thioester functionality in selected ratios to afford partial branching (Scheme 3).

Synthetic Methods

In general, the polymers of the invention can be prepared by methods described herein or by other methods known to those skilled in the art. Exemplary preparations of the polymers of the invention are described below.

The following generic schemes and examples illustrate how to prepare the polymers of the present disclosure. For example, stoichiometry of the reagents may be varied in order to obtain the desired monomer or polymer.

As shown in pathway A in Scheme 4, above, the intermediate phosphate triester 1.3 can be prepared, e.g., by treating p-nitrophenol with phosphoryl chloride (phosphorus oxychloride) in the presence of a suitable base such as triethylamine to produce intermediate phosphorochloridate 1.1, followed by treatment with S-(2-hydroxyethyl) ethanethioate in the presence of a suitable base such as triethylamine. Alternatively, as shown in pathway B, S-(2-hydroxyethyl) ethanethioate can first be treated with phosphoryl chloride in the presence of, e.g., a suitable base such as triethylamine to produce intermediate 1.2, followed by treatment with p-nitrophenol in the presence of a suitable base such as triethylamine to yield phosphate triester 1.3.

As shown in Scheme 5, above, the intermediate phosphate triester 1.3 can be converted to the moiety of Formula (I) by treatment with, e.g., an appropriately substituted diol 1.4 in the presence of (C) a suitable base such as 1,8-diazabicyclo[5.4.0]undec-7-ene in a suitable solvent or combination of solvents such as acetonitrile/dichloromethane or (D) a suitable Grignard reagent such as tert-butylmagnesium chloride in a suitable solvent such as tetrahydrofuran, followed by treatment with a suitable protic agent such as methanol.

Scheme 6, above, illustrates alternative routes to the moiety of Formula (I). For example, treatment of S-(2-hydroxyethyl) ethanethioate with N,N′-diisopropylphosphorodiamidic chloride in the presence of a suitable base such as triethylamine in a suitable solvent such as diethyl ether yields the intermediate phosphorodiamidate 1.5. Subsequent treatment with an appropriately substituted diol 1.4 in the presence of a suitable base such as nucleophilic catalysts or combinations thereof (e.g., tetrazole and/or pyridine) yields the moiety of Formula (I). Alternatively, treatment of S-(2-hydroxyethyl) ethanethioate with 1-chloro-N,N′-diisopropylphosphanediamine in the presence of a suitable base such as triethylamine in a suitable solvent such as diethyl ether yields the intermediate alkoxyphosphanediamine 1.6. Subsequent treatment with an appropriately substituted diol 1.4 in the presence of a suitable nucleophilic catalyst such as tetrazole or pyridine followed by addition of a suitable oxidizing agent such as tert-butyl peroxide yields the moiety of Formula (I).

Scheme 7, above, illustrates another alternative route to the moiety of Formula (I). Treatment of an appropriately substituted diol 1.4 with phosphoryl chloride (phosphorus oxychloride) in a suitable solvent such as methylene chloride (dichloromethane) in the presence of a suitable base such as triethylamine yields intermediate phosphorochloridate 1.7. Subsequent treatment of 1.7 with of S-(2-hydroxyethyl) ethanethioate in a suitable solvent such as tetrahydrofuran in the presence of a suitable base such as triethylamine yields phosphate triester 1.8. Ring-opening polymerization (ROP) of 1.8 yields the moiety of Formula (I).

Specific examples of poly(phosphoester) preparation are described in the following paragraphs.

Nucleic Acids

Synthesis of Nucleic Acids

Nucleic acids according to the present invention may be synthesized according to any known methods. For example, mRNAs according to the present invention may be synthesized via in vitro transcription (IVT). Briefly, IVT is typically performed with a linear or circular DNA template containing a promoter, a pool of ribonucleotide triphosphates, a buffer system that may include DTT and magnesium ions, and an appropriate RNA polymerase (e.g., T3, T7, mutated T7 or SP6 RNA polymerase), DNAse I, pyrophosphatase, and/or RNAse inhibitor. The exact conditions will vary according to the specific application.

In some embodiments, for the preparation of mRNA according to the invention, a DNA template is transcribed in vitro. A suitable DNA template typically has a promoter, for example a T3, T7, mutated T7 or SP6 promoter, for in vitro transcription, followed by desired nucleotide sequence for desired mRNA and a termination signal.

Desired mRNA sequence(s) according to the invention may be determined and incorporated into a DNA template using standard methods. For example, starting from a desired amino acid sequence (e.g., an enzyme sequence), a virtual reverse translation is carried out based on the degenerated genetic code. Optimization algorithms may then be used for selection of suitable codons. Typically, the G/C content can be optimized to achieve the highest possible G/C content on one hand, taking into the best possible account the frequency of the tRNAs according to codon usage on the other hand. The optimized RNA sequence can be established and displayed, for example, with the aid of an appropriate display device and compared with the original (wild-type) sequence. A secondary structure can also be analyzed to calculate stabilizing and destabilizing properties or, respectively, regions of the RNA.

Modified Nucleic Acids

Modified nucleic acids according to the present invention may be include modifications and be prepared according to any known modifications and methods. For example, in some embodiments, mRNA according to the present invention may be synthesized as unmodified RNA or as modified RNA in accordance with known modifications and methods. Typically, RNAs are modified to enhance stability. Modifications of RNA can include, for example, modifications of the nucleotides of the RNA. A modified RNA such as modified mRNA (mmRNA) according to the invention can thus include, for example, backbone modifications, sugar modifications or base modifications. In some embodiments, mRNAs may be synthesized from naturally occurring nucleotides and/or nucleotide analogues (modified nucleotides) including, but not limited to, purines (adenine (A), guanine (G)) or pyrimidines (thymine (T), cytosine (C), uracil (U)), and as modified nucleotides analogues or derivatives of purines and pyrimidines, such as e.g. 1-methyl-adenine, 2-methyl-adenine, 2-methylthio-N-6-isopentenyl-adenine, N6-methyl-adenine, N6-isopentenyl-adenine, 2-thio-cytosine, 3-methyl-cytosine, 4-acetyl-cytosine, 5-methyl-cytosine, 2,6-diaminopurine, 1-methyl-guanine, 2-methyl-guanine, 2,2-dimethyl-guanine, 7-methyl-guanine, inosine, 1-methyl-inosine, pseudouracil (5-uracil), dihydro-uracil, 2-thio-uracil, 4-thio-uracil, 5-carboxymethylaminomethyl-2-thio-uracil, 5-(carboxyhydroxymethyl)-uracil, 5-fluoro-uracil, 5-bromo-uracil, 5-carboxymethylaminomethyl-uracil, 5-methyl-2-thio-uracil, 5-methyl-uracil, N-uracil-5-oxyacetic acid methyl ester, 5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thio-uracil, 5′-methoxycarbonylmethyl-uracil, 5-methoxy-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), 1-methyl-pseudouracil, queosine, .beta.-D-mannosyl-queosine, wybutoxosine, and phosphoramidates, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. The preparation of such analogues is known to a person skilled in the art e.g., from the U.S. Pat. Nos. 4,373,071, 4,401,796, 4,415,732, 4,458,066, 4,500,707, 4,668,777, 4,973,679, 5,047,524, 5,132,418, 5,153,319, 5,262,530 and 5,700,642, the disclosures of which are incorporated by reference in their entirety.

In some embodiments, modified RNAs such as mmRNAs may contain RNA backbone modifications. Typically, a backbone modification is a modification in which the phosphates of the backbone of the nucleotides contained in the RNA are modified chemically. Exemplary backbone modifications typically include, but are not limited to, modifications from the group consisting of methylphosphonates, methylphosphoramidates, phosphoramidates, phosphorothioates (e.g. cytidine 5′-O-(1-thiophosphate)), boranophosphates, positively charged guanidinium groups etc., which means by replacing the phosphodiester linkage by other anionic, cationic or neutral groups.

In some embodiments, modified RNAs such as mmRNAs may contain sugar modifications. A typical sugar modification is a chemical modification of the sugar of the nucleotides it contains including, but not limited to, sugar modifications chosen from the group consisting of 4′-thio-ribonucleotide (see, e.g., US Patent Application Publication No. US 2016/0031928, incorporated by reference herein), 2′-deoxy-2′-fluoro-oligoribonucleotide (2′-fluoro-2′-deoxycytidine 5′-triphosphate, 2′-fluoro-2′-deoxyuridine 5′-triphosphate), 2′-deoxy-2′-deamine-oligoribonucleotide (2′-amino-2′-deoxycytidine 5′-triphosphate, 2′-amino-2′-deoxyuridine 5′-triphosphate), 2′-O-alkyloligoribonucleotide, 2′-deoxy-2′-C-alkyloligoribonucleotide (2′-O-methylcytidine 5′-triphosphate, 2′-methyluridine 5′-triphosphate), 2′-C-alkyloligoribonucleotide, and isomers thereof (2′-aracytidine 5′-triphosphate, 2′-arauridine 5′-triphosphate), or azidotriphosphates (2′-azido-2′-deoxycytidine 5′-triphosphate, 2′-azido-2′-deoxyuridine 5′-triphosphate).

In some embodiments, modified RNAs such as mmRNAs may contain modifications of the bases of the nucleotides (base modifications). A modified nucleotide which contains a base modification is also called a base-modified nucleotide. Examples of such base-modified nucleotides include, but are not limited to, 2-amino-6-chloropurine riboside 5′-triphosphate, 2-aminoadenosine 5′-triphosphate, 2-thiocytidine 5′-triphosphate, 2-thiouridine 5′-triphosphate, 4-thiouridine 5′-triphosphate, 5-aminoallylcytidine 5′-triphosphate, 5-aminoallyluridine 5′-triphosphate, 5-bromocytidine 5′-triphosphate, 5-bromouridine 5′-triphosphate, 5-iodocytidine 5′-triphosphate, 5-iodouridine 5′-triphosphate, 5-methylcytidine 5′-triphosphate, 5-methyluridine 5′-triphosphate, 6-azacytidine 5′-triphosphate, 6-azauridine 5′-triphosphate, 6-chloropurine riboside 5′-triphosphate, 7-deazaadenosine 5′-triphosphate, 7-deazaguanosine 5′-triphosphate, 8-azaadenosine 5′-triphosphate, 8-azidoadenosine 5′-triphosphate, benzimidazole riboside 5′-triphosphate, N1-methyladenosine 5′-triphosphate, N1-methylguanosine 5′-triphosphate, N6-methyladenosine 5′-triphosphate, 06-methylguanosine 5′-triphosphate, pseudouridine 5′-triphosphate, puromycin 5′-triphosphate or xanthosine 5′-triphosphate.

Typically, mRNA synthesis includes the addition of a “cap” on the N-terminal (5′) end, and a “tail” on the C-terminal (3′) end. The presence of the cap is important in providing resistance to nucleases found in most eukaryotic cells. The presence of a “tail” serves to protect the mRNA from exonuclease degradation.

Thus, in some embodiments, mRNAs include a 5′ cap structure. A 5′ cap is typically added as follows: first, an RNA terminal phosphatase removes one of the terminal phosphate groups from the 5′ nucleotide, leaving two terminal phosphates; guanosine triphosphate (GTP) is then added to the terminal phosphates via a guanylyl transferase, producing a 5′5′5 triphosphate linkage; and the 7-nitrogen of guanine is then methylated by a methyltransferase. Examples of cap structures include, but are not limited to, m7G(5′)ppp (5′(A,G(5′)ppp(5′)A and G(5′)ppp(5′)G.

In some embodiments, mRNAs include a 3′ poly(A) tail structure. A poly-A tail on the 3′ terminus of mRNA typically includes about 10 to 300 adenosine nucleotides (SEQ ID NO: 4) (e.g., about 10 to 200 adenosine nucleotides, about 10 to 150 adenosine nucleotides, about 10 to 100 adenosine nucleotides, about 20 to 70 adenosine nucleotides, or about 20 to 60 adenosine nucleotides). In some embodiments, mRNAs include a 3′ poly(C) tail structure. A suitable poly-C tail on the 3′ terminus of mRNA typically include about 10 to 200 cytosine nucleotides (SEQ ID NO: 5) (e.g., about 10 to 150 cytosine nucleotides, about 10 to 100 cytosine nucleotides, about 20 to 70 cytosine nucleotides, about 20 to 60 cytosine nucleotides, or about 10 to 40 cytosine nucleotides). The poly-C tail may be added to the poly-A tail or may substitute the poly-A tail.

In some embodiments, mRNAs include a 5′ and/or 3′ untranslated region. In some embodiments, a 5′ untranslated region includes one or more elements that affect an mRNA's stability or translation, for example, an iron responsive element. In some embodiments, a 5′ untranslated region may be between about 50 and 500 nucleotides in length.

In some embodiments, a 3′ untranslated region includes one or more of a polyadenylation signal, a binding site for proteins that affect an mRNA's stability of location in a cell, or one or more binding sites for miRNAs. In some embodiments, a 3′ untranslated region may be between 50 and 500 nucleotides in length or longer.

Cap Structure

In some embodiments, mRNAs include a 5′ cap structure. A 5′ cap is typically added as follows: first, an RNA terminal phosphatase removes one of the terminal phosphate groups from the 5′ nucleotide, leaving two terminal phosphates; guanosine triphosphate (GTP) is then added to the terminal phosphates via a guanylyl transferase, producing a 5′5′5 triphosphate linkage; and the 7-nitrogen of guanine is then methylated by a methyltransferase. Examples of cap structures include, but are not limited to, m7G(5′)ppp (5′(A,G(5′)ppp(5′)A and G(5′)ppp(5′)G.

Naturally occurring cap structures comprise a 7-methyl guanosine that is linked via a triphosphate bridge to the 5′-end of the first transcribed nucleotide, resulting in a dinucleotide cap of m⁷G(5′)ppp(5′)N, where N is any nucleoside. In vivo, the cap is added enzymatically. The cap is added in the nucleus and is catalyzed by the enzyme guanylyl transferase. The addition of the cap to the 5′ terminal end of RNA occurs immediately after initiation of transcription. The terminal nucleoside is typically a guanosine, and is in the reverse orientation to all the other nucleotides, i.e., G(5′)ppp(5′)GpNpNp.

A common cap for mRNA produced by in vitro transcription is m⁷G(5′)ppp(5′)G, which has been used as the dinucleotide cap in transcription with T7 or SP6 RNA polymerase in vitro to obtain RNAs having a cap structure in their 5′-termini. The prevailing method for the in vitro synthesis of capped mRNA employs a pre-formed dinucleotide of the form m⁷G(5′)ppp(5′)G (“m⁷GpppG”) as an initiator of transcription.

To date, a usual form of a synthetic dinucleotide cap used in in vitro translation experiments is the Anti-Reverse Cap Analog (“ARCA”) or modified ARCA, which is generally a modified cap analog in which the 2′ or 3′ OH group is replaced with —OCH₃.

Additional cap analogs include, but are not limited to, a chemical structures selected from the group consisting of m⁷GpppG, m⁷GpppA, m⁷GpppC; unmethylated cap analogs (e.g., GpppG); dimethylated cap analog (e.g., m^(2,7)GpppG), trimethylated cap analog (e.g., m^(2,2,7)GpppG), dimethylated symmetrical cap analogs (e.g., m⁷Gpppm⁷G), or anti reverse cap analogs (e.g., ARCA; m^(7,2′Ome)GpppG, m^(7,2′d)GpppG, m^(7,3′Ome)GpppG, m^(7,3′d)GpppG and their tetra phosphate derivatives) (see, e.g., Jemielity, J. et al., “Novel ‘anti-reverse’ cap analogs with superior translational properties”, RNA, 9: 1108-1122 (2003)).

In some embodiments, a suitable cap is a 7-methyl guanylate (“m⁷G”) linked via a triphosphate bridge to the 5′-end of the first transcribed nucleotide, resulting in m⁷G(5′)ppp(5′)N, where N is any nucleoside. A preferred embodiment of a m⁷G cap utilized in embodiments of the invention is m⁷G(5′)ppp(5′)G.

In some embodiments, the cap is a Cap0 structure. Cap0 structures lack a 2′-O-methyl residue of the ribose attached to bases 1 and 2. In some embodiments, the cap is a Cap1 structure. Cap1 structures have a 2′-O-methyl residue at base 2. In some embodiments, the cap is a Cap2 structure. Cap2 structures have a 2′-O-methyl residue attached to both bases 2 and 3.

A variety of m⁷G cap analogs are known in the art, many of which are commercially available. These include the m⁷GpppG described above, as well as the ARCA 3′-OCH₃ and 2′-OCH₃ cap analogs (Jemielity, J. et al., RNA, 9: 1108-1122 (2003)). Additional cap analogs for use in embodiments of the invention include N7-benzylated dinucleoside tetraphosphate analogs (described in Grudzien, E. et al., RNA, 10: 1479-1487 (2004)), phosphorothioate cap analogs (described in Grudzien-Nogalska, E., et al., RNA, 13: 1745-1755 (2007)), and cap analogs (including biotinylated cap analogs) described in U.S. Pat. Nos. 8,093,367 and 8,304,529, incorporated by reference herein.

Tail Structure

Typically, the presence of a “tail” serves to protect the mRNA from exonuclease degradation. The poly A tail is thought to stabilize natural messengers and synthetic sense RNA. Therefore, in certain embodiments a long poly A tail can be added to an mRNA molecule thus rendering the RNA more stable. Poly A tails can be added using a variety of art-recognized techniques. For example, long poly A tails can be added to synthetic or in vitro transcribed RNA using poly A polymerase (Yokoe, et al. Nature Biotechnology. 1996; 14: 1252-1256). A transcription vector can also encode long poly A tails. In addition, poly A tails can be added by transcription directly from PCR products. Poly A may also be ligated to the 3′ end of a sense RNA with RNA ligase (see, e.g., Molecular Cloning A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press: 1991 edition)).

In some embodiments, mRNAs include a 3′ poly(A) tail structure. Typically, the length of the poly A tail can be at least about 10, 50, 100, 200, 300, 400 at least 500 nucleotides. In some embodiments, a poly-A tail on the 3′ terminus of mRNA typically includes about 10 to 300 adenosine nucleotides (e.g., about 10 to 200 adenosine nucleotides, about 10 to 150 adenosine nucleotides, about 10 to 100 adenosine nucleotides, about 20 to 70 adenosine nucleotides, or about 20 to 60 adenosine nucleotides). In some embodiments, mRNAs include a 3′ poly(C) tail structure. A suitable poly-C tail on the 3′ terminus of mRNA typically include about 10 to 200 cytosine nucleotides (e.g., about 10 to 150 cytosine nucleotides, about 10 to 100 cytosine nucleotides, about 20 to 70 cytosine nucleotides, about 20 to 60 cytosine nucleotides, or about 10 to 40 cytosine nucleotides). The poly-C tail may be added to the poly-A tail or may substitute the poly-A tail.

In some embodiments, the length of the poly A or poly C tail is adjusted to control the stability of a modified sense mRNA molecule of the invention and, thus, the transcription of protein. For example, since the length of the poly A tail can influence the half-life of a sense mRNA molecule, the length of the poly A tail can be adjusted to modify the level of resistance of the mRNA to nucleases and thereby control the time course of polynucleotide expression and/or polypeptide production in a target cell.

5′ and 3′ Untranslated Region

In some embodiments, mRNAs include a 5′ and/or 3′ untranslated region. In some embodiments, a 5′ untranslated region includes one or more elements that affect an mRNA's stability or translation, for example, an iron responsive element. In some embodiments, a 5′ untranslated region may be between about 50 and 500 nucleotides in length.

In some embodiments, a 3′ untranslated region includes one or more of a polyadenylation signal, a binding site for proteins that affect an mRNA's stability of location in a cell, or one or more binding sites for miRNAs. In some embodiments, a 3′ untranslated region may be between 50 and 500 nucleotides in length or longer.

Exemplary 3′ and/or 5′ UTR sequences can be derived from mRNA molecules which are stable (e.g., globin, actin, GAPDH, tubulin, histone, or citric acid cycle enzymes) to increase the stability of the sense mRNA molecule. For example, a 5′ UTR sequence may include a partial sequence of a CMV immediate-early 1 (IE1) gene, or a fragment thereof to improve the nuclease resistance and/or improve the half-life of the polynucleotide. Also contemplated is the inclusion of a sequence encoding human growth hormone (hGH), or a fragment thereof to the 3′ end or untranslated region of the polynucleotide (e.g., mRNA) to further stabilize the polynucleotide. Generally, these modifications improve the stability and/or pharmacokinetic properties (e.g., half-life) of the polynucleotide relative to their unmodified counterparts, and include, for example modifications made to improve such polynucleotides' resistance to in vivo nuclease digestion.

Pharmaceutical Formulations of Polymers and Nucleic Acids

Among other things, the present invention provides a pharmaceutical formulation comprising a provided polymer and one or more nucleic acids. As described below, such a polymer-based formulation for delivering nucleic acids can provide one or more advantages in delivering nucleic acids, such as mRNA, for providing a therapeutic benefit, such as producing encoded protein (where the nucleic acid is mRNA), over existing polymers.

In one aspect, the invention features a pharmaceutical composition comprising a polymer as described herein and one or more polynucleotides.

In embodiments, a polynucleotide comprises RNA (e.g., mRNA, siRNA, snoRNA, microRNA, gRNA, crRNA and combinations thereof).

In embodiments, an RNA is mRNA.

I

mRNA encodes a peptide or polypeptide for use in the delivery to or treatment of the lung of a subject or a lung cell. In embodiments, an mRNA encodes cystic fibrosis transmembrane conductance regulator (CFTR) protein.

mRNA encodes a peptide or polypeptide for use in the delivery to or treatment of the liver of a subject or a liver cell. In embodiments, an mRNA encodes ornithine transcarbamylase (OTC) protein.

In embodiments, an mRNA encodes a peptide or polypeptide for use in vaccine. In embodiments, an mRNA encodes an antigen.

In general, the polymer and nucleic acids such as mRNA are first mixed in a solution, e.g., an aqueous solution. Typically, the polymer and nucleic acids such as mRNA form a complex when mixed. In some embodiments, the polymer and nucleic acids are complexed in a form of nanoparticles.

Various sized nanoparticles may be formed according to the present invention. In various embodiments, a nanoparticle is characterized with an average diameter based on volume diameter using dynamic light scattering. In some embodiments, nanoparticles have an average diameter between about 50 nm and about 250 nm. In some embodiments, nanoparticles have an average diameter between about 50 nm and about 200 nm. In some embodiments, nanoparticles have an average diameter between about 75 nm and about 225 nm. In some embodiments, nanoparticles have an average diameter between about 100 nm and about 200 nm. In some embodiments, nanoparticles have an average diameter between about 125 nm and about 175 nm. In some embodiments, nanoparticles have an average diameter between about 140 nm and about 160 nm. In some embodiments, nanoparticles have an average diameter between about 50 nm and about 150 nm. In some embodiments, nanoparticles have an average diameter between about 75 nm and about 125 nm. In some embodiments, nanoparticles have an average diameter between about 50 nm and about 250 nm. In some embodiments, nanoparticles have an average diameter between about 90 nm and about 110 nm. In some embodiments, nanoparticles have an average diameter between about 150 nm and about 250 nm. In some embodiments, nanoparticles have an average diameter between about 175 nm and about 225 nm. In some embodiments, nanoparticles have an average diameter between about 190 nm and about 210 nm.

Nanoparticles may have a range of sizes in a mixture. Typically, the nanoparticles have a polydispersity index (PDI) of less than about 0.5 (e.g., less than about 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, or 0.1). In some embodiment, the nanoparticles have a PDI between about 0.1 and about 0.4. In some embodiments nanoparticles have a PDI between about 0.1 and about 0.4. In some embodiments nanoparticles have a PDI between about 0.1 and about 0.2. In some embodiments nanoparticles have a PDI between about 0.2 and about 0.3. In some embodiments, nanoparticles have a PDI between about 0.3 and about 0.4.

Nanoparticles may have a range of surface charges as measured through zeta potential measurements. In some embodiments, nanoparticles have a zeta potential between 0 and about 50 mV. In some embodiments nanoparticles have a zeta potential between about 5 and about 45 mV. In some embodiments nanoparticles have a zeta potential between about 10 and about 40 mV. In some embodiments nanoparticles have a zeta potential between about 10 and about 30 mV. In some embodiments nanoparticles have a zeta potential between about 20 and about 30 mV.

Pharmaceutical Formulations and Therapeutic Uses

Pharmaceutical formulations containing provided polymer and nucleic acids provided by the present invention may be used for various therapeutic purposes. Provided polymer-based formulations for delivering nucleic acids can provide one or more advantages in delivering nucleic acids, such as mRNA, for providing a therapeutic benefit, such as producing encoded protein (where the nucleic acid is mRNA), over existing polymers.

Provided polymers are biodegradable. Biodegradable polymers can provide a higher level of tolerability than other polymeric and lipidic delivery vehicles.

Provided polymers are ionizable. The ionizability can be engendered by an amino functionality in the polymer backbone (e.g., R is N) and/or a protonatable group tethered from the backbone. The protonatable group will have at least one 1° amino, 2° amino, 3° amino or imidazolyl functionality. Such ionizable groups can provide a buffering effect and act as a “proton sponge” allowing for enhancement of endosomal release.

Provided polymers incorporate an S-acycl-2-thioethyl (“SATE”) protecting group on the phosphate groups within the backbone. Such protection removes the negative charge from the phosphate groups, allowing for facile complexation of the ionizable positively charged polymer with nucleic acids. Upon cellular entry, the SATE protecting groups are biolabile and can be cleaved enzymatically by carboxylesterase. Such cleavage and results in a negatively charged phosphate moiety across the backbone of polymers. This, in effect, neutralizes the polymer charge and thus removes the electrostatic ability of the polymer to bind nucleic acids, e.g., mRNA. This can result in efficient release of the payload nucleic acid.

To facilitate delivery of nucleic acids in vivo, polymer/nucleic acids can be formulated in combination with one or more additional pharmaceutical carriers, targeting ligands or stabilizing reagents. Techniques for formulation and administration of drugs may be found in “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., latest edition.

Suitable routes of administration include, for example, oral, rectal, vaginal, transmucosal, pulmonary including intratracheal or inhaled, or intestinal administration; parenteral delivery, including intradermal, transdermal (topical), intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, or intranasal. In particular embodiments, the intramuscular administration is to a muscle selected from the group consisting of skeletal muscle, smooth muscle and cardiac muscle. In some embodiments the administration results in delivery of the nucleic acids to a muscle cell. In some embodiments the administration results in delivery of the nucleic acids to a hepatocyte (i.e., liver cell).

Alternatively or additionally, pharmaceutical formulations of the invention may be administered in a local rather than systemic manner, for example, via injection of the pharmaceutical formulation directly into a targeted tissue, preferably in a sustained release formulation. Local delivery can be affected in various ways, depending on the tissue to be targeted. For example, aerosols containing compositions of the present invention can be inhaled (for nasal, tracheal, or bronchial delivery); compositions of the present invention can be injected into the site of injury, disease manifestation, or pain, for example; compositions can be provided in lozenges for oral, tracheal, or esophageal application; can be supplied in liquid, tablet or capsule form for administration to the stomach or intestines, can be supplied in suppository form for rectal or vaginal application; or can even be delivered to the eye by use of creams, drops, or even injection.

The present invention provides methods for producing a composition enriched with full-length mRNA molecules which are greater than 500 nucleotides in length and encoding for a peptide or polypeptide of interest. The present invention also provides methods for producing a therapeutic composition enriched with full-length mRNA molecules encoding a peptide or polypeptide of interest for use in the delivery to or treatment of a subject, e.g., a human subject or a cell of a human subject or a cell that is treated and delivered to a human subject.

Accordingly, in certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the lung of a subject or a lung cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for cystic fibrosis transmembrane conductance regulator (CFTR) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for ATP-binding cassette sub-family A member 3 protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for dynein axonemal intermediate chain 1 protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for dynein axonemal heavy chain 5 (DNAH5) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for alpha-1-antitrypsin protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for forkhead box P3 (FOXP3) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes one or more surfactant protein, e.g., one or more of surfactant A protein, surfactant B protein, surfactant C protein, and surfactant D protein.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the liver of a subject or a liver cell. Such peptides and polypeptides can include those associated with a urea cycle disorder, associated with a lysosomal storage disorder, with a glycogen storage disorder, associated with an amino acid metabolism disorder, associated with a lipid metabolism or fibrotic disorder, associated with methylmalonic acidemia, or associated with any other metabolic disorder for which delivery to or treatment of the liver or a liver cell with enriched full-length mRNA provides therapeutic benefit.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein associated with a urea cycle disorder. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for ornithine transcarbamylase (OTC) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for arginosuccinate synthetase 1 protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for carbamoyl phosphate synthetase I protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for arginosuccinate lyase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for arginase protein.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein associated with a lysosomal storage disorder. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for alpha galactosidase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for glucocerebrosidase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for iduronate-2-sulfatase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for iduronidase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for N-acetyl-alpha-D-glucosaminidase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for heparan N-sulfatase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for galactosamine-6 sulfatase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for beta-galactosidase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for lysosomal lipase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for arylsulfatase B (N-acetylgalactosamine-4-sulfatase) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for transcription factor EB (TFEB).

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein associated with a glycogen storage disorder. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for acid alpha-glucosidase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for glucose-6-phosphatase (G6PC) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for liver glycogen phosphorylase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for muscle phosphoglycerate mutase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for glycogen debranching enzyme.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein associated with amino acid metabolism. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for phenylalanine hydroxylase enzyme. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for glutaryl-CoA dehydrogenase enzyme. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for propionyl-CoA caboxylase enzyme. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for oxalase alanine-glyoxylate aminotransferase enzyme.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein associated with a lipid metabolism or fibrotic disorder. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a mTOR inhibitor. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for ATPase phospholipid transporting 8B1 (ATP8B1) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for one or more NF-kappa B inhibitors, such as one or more of I-kappa B alpha, interferon-related development regulator 1 (IFRD1), and Sirtuin 1 (SIRT1). In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for PPAR-gamma protein or an active variant.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein associated with methylmalonic acidemia. For example, in certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for methylmalonyl CoA mutase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for methylmalonyl CoA epimerase protein.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA for which delivery to or treatment of the liver can provide therapeutic benefit. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for ATP7B protein, also known as Wilson disease protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for porphobilinogen deaminase enzyme. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for one or clotting enzymes, such as Factor VIII, Factor IX, Factor VII, and Factor X. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for human hemochromatosis (HFE) protein.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the cardiovasculature of a subject or a cardiovascular cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for vascular endothelial growth factor A protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for relaxin protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for bone morphogenetic protein-9 protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for bone morphogenetic protein-2 receptor protein.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the muscle of a subject or a muscle cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for dystrophin protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for frataxin protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the cardiac muscle of a subject or a cardiac muscle cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein that modulates one or both of a potassium channel and a sodium channel in muscle tissue or in a muscle cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein that modulates a Kv7.1 channel in muscle tissue or in a muscle cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a protein that modulates a Nav1.5 channel in muscle tissue or in a muscle cell.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the nervous system of a subject or a nervous system cell. For example, in certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for survival motor neuron 1 protein. For example, in certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for survival motor neuron 2 protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for frataxin protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for ATP binding cassette subfamily D member 1 (ABCD1) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for CLN3 protein.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the blood or bone marrow of a subject or a blood or bone marrow cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for beta globin protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for Bruton's tyrosine kinase protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for one or clotting enzymes, such as Factor VIII, Factor IX, Factor VII, and Factor X.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the kidney of a subject or a kidney cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for collagen type IV alpha 5 chain (COL4A5) protein.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery to or treatment of the eye of a subject or an eye cell. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for ATP-binding cassette sub-family A member 4 (ABCA4) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for retinoschisin protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for retinal pigment epithelium-specific 65 kDa (RPE65) protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for centrosomal protein of 290 kDa (CEP290).

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes a peptide or polypeptide for use in the delivery of or treatment with a vaccine for a subject or a cell of a subject. For example, in certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from an infectious agent, such as a virus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from influenza virus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from respiratory syncytial virus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from rabies virus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from cytomegalovirus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from rotavirus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from a hepatitis virus, such as hepatitis A virus, hepatitis B virus, or hepatis C virus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from human papillomavirus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from a herpes simplex virus, such as herpes simplex virus 1 or herpes simplex virus 2. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from a human immunodeficiency virus, such as human immunodeficiency virus type 1 or human immunodeficiency virus type 2. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from a human metapneumovirus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from a human parainfluenza virus, such as human parainfluenza virus type 1, human parainfluenza virus type 2, or human parainfluenza virus type 3. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from malaria virus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from zika virus. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen from chikungunya virus.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen associated with a cancer of a subject or identified from a cancer cell of a subject. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen determined from a subject's own cancer cell, i.e., to provide a personalized cancer vaccine. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antigen expressed from a mutant KRAS gene.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antibody. In certain embodiments, the antibody can be a bi-specific antibody. In certain embodiments, the antibody can be part of a fusion protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antibody to OX40. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antibody to VEGF. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antibody to tissue necrosis factor alpha. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antibody to CD3. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an antibody to CD19.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an immunomodulator. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for Interleukin 12. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for Interleukin 23. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for Interleukin 36 gamma. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a constitutively active variant of one or more stimulator of interferon genes (STING) proteins.

In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an endonuclease. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for an RNA-guided DNA endonuclease protein, such as Cas 9 protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a meganuclease protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a transcription activator-like effector nuclease protein. In certain embodiments the present invention provides a method for producing a therapeutic composition enriched with full-length mRNA that encodes for a zinc finger nuclease protein.

In embodiments, exemplary therapeutic uses result from the delivery of mRNA encoding a secreted protein. Accordingly, in embodiments, the compositions and methods of the invention provide for delivery of mRNA encoding a secreted protein. In some embodiments, the compositions and methods of the invention provide for delivery of mRNA encoding one or more secreted proteins listed in Table 1; thus, compositions of the invention may comprise an mRNA encoding a protein listed in Table 1 (or a homolog thereof) along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein listed in Table 1 (or a homolog thereof) along with other components set out herein.

In some embodiments, compositions and methods of the invention provide for the delivery of one or more nucleic acids that bind to or otherwise act on an endogenous nucleic acid, such as endogenous mRNA or DNA, that alter the transcription, translation, secretion, or action of one or more proteins or genes listed in Table 1 (or a homolog thereof). For example, various RNAi nucleic acids bind to endogenous mRNA, miRNA, or other RNAs, while guide RNA (gRNA) and CRISPR RNA (crRNA) respectively bind to DNA and act on DNA in the nucleus. Accordingly, in embodiments, the compositions and methods of the invention provide for delivery of nucleic acids including one or more of RNAi nucleic acids, gRNA and crRNA that interfere with the transcription, translation, secretion, or action of one or more genes or proteins listed in Table 1 (or homologs thereof) along with other components set out herein.

TABLE 1 Secreted Proteins Uniprot ID Protein Name Gene Name A1E959 Odontogenic ameloblast-associated protein ODAM A1KZ92 Peroxidasin-like protein PXDNL A1L453 Serine protease 38 PRSS38 A1L4H1 Soluble scavenger receptor cysteine-rich domain- SSC5D containing protein SSC5D A2RUU4 Colipase-like protein 1 CLPSL1 A2VDF0 Fucose mutarotase FUOM A2VEC9 SCO-spondin SSPO A3KMH1 von Willebrand factor A domain-containing VWA8 protein 8 A4D0S4 Laminin subunit beta-4 LAMB4 A4D1T9 Probable inactive serine protease 37 PRSS37 A5D8T8 C-type lectin domain family 18 member A CLEC18A A6NC86 phospholipase A2 inhibitor and Ly6/PLAUR PINLYP domain-containing protein A6NCI4 von Willebrand factor A domain-containing VWA3A protein 3A A6ND01 Probable folate receptor delta FOLR4 A6NDD2 Beta-defensin 108B-like A6NE02 BTB/POZ domain-containing protein 17 BTBD17 A6NEF6 Growth hormone 1 GH1 A6NF02 NPIP-like protein LOC730153 A6NFB4 HCG1749481, isoform CRA_k CSH1 A6NFZ4 Protein FAM24A FAM24A A6NG13 Glycosyltransferase 54 domain-containing protein A6NGN9 IgLON family member 5 IGLON5 A6NHN0 Otolin-1 OTOL1 A6NHN6 Nuclear pore complex-interacting protein-like 2 NPIPL2 A6NI73 Leukocyte immunoglobulin-like receptor LILRA5 subfamily A member 5 A6NIT4 Chorionic somatomammotropin hormone 2 CSH2 isoform 2 A6NJ69 IgA-inducing protein homolog IGIP A6NKQ9 Choriogonadotropin subunit beta variant 1 CGB1 A6NMZ7 Collagen alpha-6(VI) chain COL6A6 A6NNS2 Dehydrogenase/reductase SDR family member 7C DHRS7C A6XGL2 Insulin A chain INS A8K0G1 Protein Wnt WNT7B A8K2U0 Alpha-2-macroglobulin-like protein 1 A2ML1 A8K7I4 Calcium-activated chloride channel regulator 1 CLCA1 A8MTL9 Serpin-like protein HMSD HMSD A8MV23 Serpin E3 SERPINE3 A8MZH6 Oocyte-secreted protein 1 homolog OOSP1 A8TX70 Collagen alpha-5(VI) chain COL6A5 B0ZBE8 Natriuretic peptide NPPA B1A4G9 Somatotropin GH1 B1A4H2 HCG1749481, isoform CRA_d CSH1 B1A4H9 Chorionic somatomammotropin hormone CSH2 B1AJZ6 Protein Wnt WNT4 B1AKI9 Isthmin-1 ISM1 B2RNN3 Complement C1q and tumor necrosis factor- C1QTNF9B related protein 9B B2RUY7 von Willebrand factor C domain-containing VWC2L protein 2-like B3GLJ2 Prostate and testis expressed protein 3 PATE3 B4DI03 SEC11-like 3 (S. cerevisiae), isoform CRA_a SEC11L3 B4DJF9 Protein Wnt WNT4 B4DUL4 SEC11-like 1 (S. cerevisiae), isoform CRA_d SEC11L1 B5MCC8 Protein Wnt WNT10B B8A595 Protein Wnt WNT7B B8A597 Protein Wnt WNT7B B8A598 Protein Wnt WNT7B B9A064 Immunoglobulin lambda-like polypeptide 5 IGLL5 C9J3H3 Protein Wnt WNT10B C9J8I8 Protein Wnt WNT5A C9JAF2 Insulin-like growth factor II Ala-25 Del IGF2 C9JCI2 Protein Wnt WNT10B C9JL84 HERV-H LTR-associating protein 1 HHLA1 C9JNR5 Insulin A chain INS C9JUI2 Protein Wnt WNT2 D6RF47 Protein Wnt WNT8A D6RF94 Protein Wnt WNT8A E2RYF7 Protein PBMUCL2 HCG22 E5RFR1 PENK(114-133) PENK E7EML9 Serine protease 44 PRSS44 E7EPC3 Protein Wnt WNT9B E7EVP0 Nociceptin PNOC E9PD02 Insulin-like growth factor I IGF1 E9PH60 Protein Wnt WNT16 E9PJL6 Protein Wnt WNT11 F5GYM2 Protein Wnt WNT5B F5H034 Protein Wnt WNT5B F5H364 Protein Wnt WNT5B F5H7Q6 Protein Wnt WNT5B F8WCM5 Protein INS-IGF2 INS-IGF2 F8WDR1 Protein Wnt WNT2 H0Y663 Protein Wnt WNT4 H0YK72 Signal peptidase complex catalytic subunit SEC11A SEC11A H0YK83 Signal peptidase complex catalytic subunit SEC11A SEC11A H0YM39 Chorionic somatomammotropin hormone CSH2 H0YMT7 Chorionic somatomammotropin hormone CSH1 H0YN17 Chorionic somatomammotropin hormone CSH2 H0YNA5 Signal peptidase complex catalytic subunit SEC11A SEC11A H0YNG3 Signal peptidase complex catalytic subunit SEC11A SEC11A H0YNX5 Signal peptidase complex catalytic subunit SEC11A SEC11A H7BZB8 Protein Wnt WNT10A H9KV56 Choriogonadotropin subunit beta variant 2 CGB2 I3L0L8 Protein Wnt WNT9B J3KNZ1 Choriogonadotropin subunit beta variant 1 CGB1 J3KP00 Choriogonadotropin subunit beta CGB7 J3QT02 Choriogonadotropin subunit beta variant 1 CGB1 O00175 C-C motif chemokine 24 CCL24 O00182 Galectin-9 LGALS9 O00187 Mannan-binding lectin serine protease 2 MASP2 O00230 Cortistatin CORT O00253 Agouti-related protein AGRP O00270 12-(S)-hydroxy-5,8,10,14-eicosatetraenoic acid GPR31 receptor O00292 Left-right determination factor 2 LEFTY2 O00294 Tubby-related protein 1 TULP1 O00295 Tubby-related protein 2 TULP2 O00300 Tumor necrosis factor receptor superfamily TNFRSF11B member 11B O00339 Matrilin-2 MATN2 O00391 Sulfhydryl oxidase 1 QSOX1 O00468 Agrin AGRN O00515 Ladinin-1 LAD1 O00533 Processed neural cell adhesion molecule L1-like CHL1 protein O00584 Ribonuclease T2 RNASET2 O00585 C-C motif chemokine 21 CCL21 O00602 Ficolin-1 FCN1 O00622 Protein CYR61 CYR61 O00626 MDC(5-69) CCL22 O00634 Netrin-3 NTN3 O00744 Protein Wnt-10b WNT10B O00755 Protein Wnt-7a WNT7A O14498 Immunoglobulin superfamily containing leucine- ISLR rich repeat protein O14511 Pro-neuregulin-2, membrane-bound isoform NRG2 O14594 Neurocan core protein NCAN O14625 C-X-C motif chemokine 11 CXCL11 O14638 Ectonucleotide ENPP3 pyrophosphatase/phosphodiesterase family member 3 O14656 Torsin-1A TOR1A O14657 Torsin-1B TOR1B O14786 Neuropilin-1 NRP1 O14788 Tumor necrosis factor ligand superfamily member TNFSF11 11, membrane form O14791 Apolipoprotein L1 APOL1 O14793 Growth/differentiation factor 8 MSTN O14904 Protein Wnt-9a WNT9A O14905 Protein Wnt-9b WNT9B O14944 Proepiregulin EREG O14960 Leukocyte cell-derived chemotaxin-2 LECT2 O15018 Processed PDZ domain-containing protein 2 PDZD2 O15041 Semaphorin-3E SEMA3E O15072 A disintegrin and metalloproteinase with ADAMTS3 thrombospondin motifs 3 O15123 Angiopoietin-2 ANGPT2 O15130 Neuropeptide FF NPFF O15197 Ephrin type-B receptor 6 EPHB6 O15204 ADAM DEC 1 ADAMDEC1 O15230 Laminin subunit alpha-5 LAMA5 O15232 Matrilin-3 MATN3 O15240 Neuroendocrine regulatory peptide-1 VGF O15263 Beta-defensin 4A DEFB4A O15335 Chondroadherin CHAD O15393 Transmembrane protease serine 2 catalytic chain TMPRSS2 O15444 C-C motif chemokine 25 CCL25 O15467 C-C motif chemokine 16 CCL16 O15496 Group 10 secretory phospholipase A2 PLA2G10 O15520 Fibroblast growth factor 10 FGF10 O15537 Retinoschisin RS1 O43157 Plexin-B1 PLXNB1 O43184 Disintegrin and metalloproteinase domain- ADAM12 containing protein 12 O43240 Kallikrein-10 KLK10 O43278 Kunitz-type protease inhibitor 1 SPINT1 O43320 Fibroblast growth factor 16 FGF16 O43323 Desert hedgehog protein C-product DHH O43405 Cochlin COCH O43508 Tumor necrosis factor ligand superfamily member TNFSF12 12, membrane form O43555 Progonadoliberin-2 GNRH2 O43557 Tumor necrosis factor ligand superfamily member TNFSF14 14, soluble form O43692 Peptidase inhibitor 15 PI15 O43699 Sialic acid-binding Ig-like lectin 6 SIGLEC6 O43820 Hyaluronidase-3 HYAL3 O43827 Angiopoietin-related protein 7 ANGPTL7 O43852 Calumenin CALU O43854 EGF-like repeat and discoidin I-like domain- EDIL3 containing protein 3 O43866 CD5 antigen-like CD5L O43897 Tolloid-like protein 1 TLL1 O43915 Vascular endothelial growth factor D FIGF O43927 C-X-C motif chemokine 13 CXCL13 O60218 Aldo-keto reductase family 1 member B10 AKR1B10 O60235 Transmembrane protease serine 11D TMPRSS11D O60258 Fibroblast growth factor 17 FGF17 O60259 Kallikrein-8 KLK8 O60383 Growth/differentiation factor 9 GDF9 O60469 Down syndrome cell adhesion molecule DSCAM O60542 Persephin PSPN O60565 Gremlin-1 GREM1 O60575 Serine protease inhibitor Kazal-type 4 SPINK4 O60676 Cystatin-8 CST8 O60687 Sushi repeat-containing protein SRPX2 SRPX2 O60844 Zymogen granule membrane protein 16 ZG16 O60882 Matrix metalloproteinase-20 MMP20 O60938 Keratocan KERA O75015 Low affinity immunoglobulin gamma Fc region FCGR3B receptor III-B O75077 Disintegrin and metalloproteinase domain- ADAM23 containing protein 23 O75093 Slit homolog 1 protein SLIT1 O75094 Slit homolog 3 protein SLIT3 O75095 Multiple epidermal growth factor-like domains MEGF6 protein 6 O75173 A disintegrin and metalloproteinase with ADAMTS4 thrombospondin motifs 4 O75200 Nuclear pore complex-interacting protein-like 1 NPIPL1 O75339 Cartilage intermediate layer protein 1 C1 CILP O75354 Ectonucleoside triphosphate diphosphohydrolase 6 ENTPD6 O75386 Tubby-related protein 3 TULP3 O75398 Deformed epidermal autoregulatory factor 1 DEAF1 homolog O75443 Alpha-tectorin TECTA O75445 Usherin USH2A O75462 Cytokine receptor-like factor 1 CRLF1 O75487 Glypican-4 GPC4 O75493 Carbonic anhydrase-related protein 11 CA11 O75594 Peptidoglycan recognition protein 1 PGLYRP1 O75596 C-type lectin domain family 3 member A CLEC3A O75610 Left-right determination factor 1 LEFTY1 O75629 Protein CREG1 CREG1 O75636 Ficolin-3 FCN3 O75711 Scrapie-responsive protein 1 SCRG1 O75715 Epididymal secretory glutathione peroxidase GPX5 O75718 Cartilage-associated protein CRTAP O75829 Chondrosurfactant protein LECT1 O75830 Serpin I2 SERPINI2 O75882 Attractin ATRN O75888 Tumor necrosis factor ligand superfamily member TNFSF13 13 O75900 Matrix metalloproteinase-23 MMP23A O75951 Lysozyme-like protein 6 LYZL6 O75973 C1q-related factor C1QL1 O76038 Secretagogin SCGN O76061 Stanniocalcin-2 STC2 O76076 WNT1-inducible-signaling pathway protein 2 WISP2 O76093 Fibroblast growth factor 18 FGF18 O76096 Cystatin-F CST7 O94769 Extracellular matrix protein 2 ECM2 O94813 Slit homolog 2 protein C-product SLIT2 O94907 Dickkopf-related protein 1 DKK1 O94919 Endonuclease domain-containing 1 protein ENDOD1 O94964 N-terminal form SOGA1 O95025 Semaphorin-3D SEMA3D O95084 Serine protease 23 PRSS23 O95150 Tumor necrosis factor ligand superfamily member TNFSF15 15 O95156 Neurexophilin-2 NXPH2 O95157 Neurexophilin-3 NXPH3 O95158 Neurexophilin-4 NXPH4 O95388 WNT1-inducible-signaling pathway protein 1 WISP1 O95389 WNT1-inducible-signaling pathway protein 3 WISP3 O95390 Growth/differentiation factor 11 GDF11 O95393 Bone morphogenetic protein 10 BMP 10 O95399 Urotensin-2 UTS2 O95407 Tumor necrosis factor receptor superfamily TNFRSF6B member 6B O95428 Papilin PAPLN O95445 Apolipoprotein M APOM O95450 A disintegrin and metalloproteinase with ADAMTS2 thrombospondin motifs 2 O95460 Matrilin-4 MATN4 O95467 LHAL tetrapeptide GNAS O95631 Netrin-1 NTN1 O95633 Follistatin-related protein 3 FSTL3 O95711 Lymphocyte antigen 86 LY86 O95715 C-X-C motif chemokine 14 CXCL14 O95750 Fibroblast growth factor 19 FGF19 O95760 Interleukin-33 IL33 O95813 Cerberus CER1 O95841 Angiopoietin-related protein 1 ANGPTL1 O95897 Noelin-2 OLFM2 O95925 Eppin EPPIN O95965 Integrin beta-like protein 1 ITGBL1 O95967 EGF-containing fibulin-like extracellular matrix EFEMP2 protein 2 O95968 Secretoglobin family 1D member 1 SCGB1D1 O95969 Secretoglobin family 1D member 2 SCGB1D2 O95970 Leucine-rich glioma-inactivated protein 1 LGI1 O95972 Bone morphogenetic protein 15 BMP15 O95994 Anterior gradient protein 2 homolog AGR2 O95998 Interleukin-18-binding protein IL18BP O96009 Napsin-A NAPSA O96014 Protein Wnt-11 WNT11 P00450 Ceruloplasmin CP P00451 Factor Villa light chain F8 P00488 Coagulation factor XIII A chain F13A1 P00533 Epidermal growth factor receptor EGFR P00709 Alpha-lactalbumin LALBA P00734 Prothrombin F2 P00738 Haptoglobin beta chain HP P00739 Haptoglobin-related protein HPR P00740 Coagulation factor IXa heavy chain F9 P00742 Factor X heavy chain F10 P00746 Complement factor D CFD P00747 Plasmin light chain B PLG P00748 Coagulation factor XIIa light chain F12 P00749 Urokinase-type plasminogen activator long PLAU chain A P00750 Tissue-type plasminogen activator PLAT P00751 Complement factor B Ba fragment CFB P00797 Renin REN P00973 2′-5′-oligoadenylate synthase 1 OAS1 P00995 Pancreatic secretory trypsin inhibitor SPINK1 P01008 Antithrombin-III SERPINC1 P01009 Alpha-1-antitrypsin SERPINA1 P01011 Alpha-1-antichymotrypsin His-Pro-less SERPINA3 P01019 Angiotensin-1 AGT P01023 Alpha-2-macroglobulin A2M P01024 Acylation stimulating protein C3 P01031 Complement C5 beta chain C5 P01033 Metalloproteinase inhibitor 1 TIMP1 P01034 Cystatin-C CST3 P01036 Cystatin-S CST4 P01037 Cystatin-SN CST1 P01042 Kininogen-1 light chain KNG1 P01127 Platelet-derived growth factor subunit B PDGFB P01135 Transforming growth factor alpha TGFA P01137 Transforming growth factor beta-1 TGFB1 P01138 Beta-nerve growth factor NGF P01148 Gonadoliberin-1 GNRH1 P01160 Atrial natriuretic factor NPPA P01178 Oxytocin OXT P01185 Vasopressin-neurophysin 2-copeptin AVP P01189 Corticotropin POMC P01210 PENK(237-258) PENK P01213 Alpha-neoendorphin PDYN P01215 Glycoprotein hormones alpha chain CGA P01222 Thyrotropin subunit beta TSHB P01225 Follitropin subunit beta FSHB P01229 Lutropin subunit beta LHB P01233 Choriogonadotropin subunit beta CGB8 P01236 Prolactin PRL P01241 Somatotropin GH1 P01242 Growth hormone variant GH2 P01243 Chorionic somatomammotropin hormone CSH2 P01258 Katacalcin CALCA P01266 Thyroglobulin TG P01270 Parathyroid hormone PTH P01275 Glucagon GCG P01282 Intestinal peptide PHM-27 VIP P01286 Somatoliberin GHRH P01298 Pancreatic prohormone PPY P01303 C-flanking peptide of NPY NPY P01308 Insulin INS P01344 Insulin-like growth factor II IGF2 P01350 Big gastrin GAST P01374 Lymphotoxin-alpha LTA P01375 C-domain 1 TNF P01562 Interferon alpha-1/13 IFNA1 P01563 Interferon alpha-2 IFNA2 P01566 Interferon alpha-10 IFNA10 P01567 Interferon alpha-7 IFNA7 P01568 Interferon alpha-21 IFNA21 P01569 Interferon alpha-5 IFNA5 P01570 Interferon alpha-14 IFNA14 P01571 Interferon alpha-17 IFNA17 P01574 Interferon beta IFNB1 P01579 Interferon gamma IFNG P01583 Interleukin-1 alpha IL1A P01584 Interleukin-1 beta IL1B P01588 Erythropoietin EPO P01591 Immunoglobulin J chain IGJ P01732 T-cell surface glycoprotein CD8 alpha chain CD8A P01833 Polymeric immunoglobulin receptor PIGR P01857 Ig gamma-1 chain C region IGHG1 P01859 Ig gamma-2 chain C region IGHG2 P01860 Ig gamma-3 chain C region IGHG3 P01861 Ig gamma-4 chain C region IGHG4 P01871 Ig mu chain C region IGHM P01880 Ig delta chain C region IGHD P02452 Collagen alpha-1(I) chain COL1A1 P02458 Chondrocalcin COL2A1 P02461 Collagen alpha-1(III) chain COL3A1 P02462 Collagen alpha-1(IV) chain COL4A1 P02647 Apolipoprotein A-I APOA1 P02649 Apolipoprotein E APOE P02652 Apolipoprotein A-II APOA2 P02654 Apolipoprotein C-I APOC1 P02655 Apolipoprotein C-II APOC2 P02656 Apolipoprotein C-III APOC3 P02671 Fibrinogen alpha chain FGA P02675 Fibrinopeptide B FGB P02679 Fibrinogen gamma chain FGG P02741 C-reactive protein CRP P02743 Serum amyloid P-component(1-203) APCS P02745 Complement C1q subcomponent subunit A C1QA P02746 Complement C1q subcomponent subunit B C1QB P02747 Complement C1q subcomponent subunit C C1QC P02748 Complement component C9b C9 P02749 Beta-2-glycoprotein 1 APOH P02750 Leucine-rich alpha-2-glycoprotein LRG1 P02751 Ugl-Y2 FN1 P02753 Retinol-binding protein 4 RBP4 P02760 Trypstatin AMBP P02763 Alpha-1-acid glycoprotein 1 ORM1 P02765 Alpha-2-HS-glycoprotein chain A AHSG P02766 Transthyretin TTR P02768 Serum albumin ALB P02771 Alpha-fetoprotein AFP P02774 Vitamin D-binding protein GC P02775 Connective tissue-activating peptide III PPBP P02776 Platelet factor 4 PF4 P02778 CXCL10(1-73) CXCL10 P02786 Transferrin receptor protein 1 TFRC P02787 Serotransferrin TF P02788 Lactoferroxin-C LTF P02790 Hemopexin HPX P02808 Statherin STATH P02810 Salivary acidic proline-rich phosphoprotein 1/2 PRH2 P02812 Basic salivary proline-rich protein 2 PRB2 P02814 Peptide D1A SMR3B P02818 Osteocalcin BGLAP P03950 Angiogenin ANG P03951 Coagulation factor XIa heavy chain F11 P03952 Plasma kallikrein KLKB1 P03956 27 kDa interstitial collagenase MMP1 P03971 Muellerian-inhibiting factor AMH P03973 Antileukoproteinase SLPI P04003 C4b-binding protein alpha chain C4BPA P04004 Somatomedin-B VTN P04054 Phospholipase A2 PLA2G1B P04085 Platelet-derived growth factor subunit A PDGFA P04090 Relaxin A chain RLN2 P04114 Apolipoprotein B-100 APOB P04118 Colipase CLPS P04141 Granulocyte-macrophage colony-stimulating CSF2 factor P04155 Trefoil factor 1 TFF1 P04180 Phosphatidylcholine-sterol acyltransferase LCAT P04196 Histidine-rich glycoprotein HRG P04217 Alpha-1B-glycoprotein A1BG P04275 von Willebrand antigen 2 VWF P04278 Sex hormone-binding globulin SHBG P04279 Alpha-inhibin-31 SEMG1 P04280 Basic salivary proline-rich protein 1 PRB1 P04628 Proto-oncogene Wnt-1 WNT1 P04745 Alpha-amylase 1 AMY1A P04746 Pancreatic alpha-amylase AMY2A P04808 Prorelaxin H1 RLN1 P05000 Interferon omega-1 IFNW1 P05013 Interferon alpha-6 IFNA6 P05014 Interferon alpha-4 IFNA4 P05015 Interferon alpha-16 IFNA16 P05019 Insulin-like growth factor I IGF1 P05060 GAWK peptide CHGB P05090 Apolipoprotein D APOD P05109 Protein S100-A8 S100A8 P05111 Inhibin alpha chain INHA P05112 Interleukin-4 IL4 P05113 Interleukin-5 IL5 P05120 Plasminogen activator inhibitor 2 SERPINB2 P05121 Plasminogen activator inhibitor 1 SERPINE1 P05154 Plasma serine protease inhibitor SERPINA5 P05155 Plasma protease C1 inhibitor SERPING1 P05156 Complement factor I heavy chain CFI P05160 Coagulation factor XIII B chain F13B P05161 Ubiquitin-like protein ISG15 ISG15 P05230 Fibroblast growth factor 1 FGF1 P05231 Interleukin-6 IL6 P05305 Big endothelin-1 EDN1 P05408 C-terminal peptide SCG5 P05451 Lithostathine-1-alpha REG1A P05452 Tetranectin CLEC3B P05543 Thyroxine-binding globulin SERPINA7 P05814 Beta-casein CSN2 P05997 Collagen alpha-2(V) chain COL5A2 P06276 Cholinesterase BCHE P06307 Cholecystokinin-12 CCK P06396 Gelsolin GSN P06681 Complement C2 C2 P06702 Protein S100-A9 S100A9 P06727 Apolipoprotein A-IV APOA4 P06734 Low affinity immunoglobulin epsilon Fc receptor FCER2 soluble form P06744 Glucose-6-phosphate isomerase GPI P06850 Corticoliberin CRH P06858 Lipoprotein lipase LPL P06881 Calcitonin gene-related peptide 1 CALCA P07093 Glia-derived nexin SERPINE2 P07098 Gastric triacylglycerol lipase LIPF P07225 Vitamin K-dependent protein S PROS1 P07237 Protein disulfide-isomerase P4HB P07288 Prostate-specific antigen KLK3 P07306 Asialoglycoprotein receptor 1 ASGR1 P07355 Annexin A2 ANXA2 P07357 Complement component C8 alpha chain C8A P07358 Complement component C8 beta chain C8B P07360 Complement component C8 gamma chain C8G P07477 Alpha-trypsin chain 2 PRSS1 P07478 Trypsin-2 PRSS2 P07492 Neuromedin-C GRP P07498 Kappa-casein CSN3 P07585 Decorin DCN P07911 Uromodulin UMOD P07942 Laminin subunit beta-1 LAMB1 P07988 Pulmonary surfactant-associated protein B SFTPB P07998 Ribonuclease pancreatic RNASE1 P08118 Beta-microseminoprotein MSMB P08123 Collagen alpha-2(I) chain COL1A2 P08185 Corticosteroid-binding globulin SERPINA6 P08217 Chymotrypsin-like elastase family member 2A CELA2A P08218 Chymotrypsin-like elastase family member 2B CELA2B P08253 72 kDa type IV collagenase MMP2 P08254 Stromelysin-1 MMP3 P08294 Extracellular superoxide dismutase [Cu—Zn] SOD3 P08476 Inhibin beta A chain INHBA P08493 Matrix Gla protein MGP P08572 Collagen alpha-2(IV) chain COL4A2 P08581 Hepatocyte growth factor receptor MET P08603 Complement factor H CFH P08620 Fibroblast growth factor 4 FGF4 P08637 Low affinity immunoglobulin gamma Fc region FCGR3A receptor III-A P08697 Alpha-2-antiplasmin SERPINF2 P08700 Interleukin-3 IL3 P08709 Coagulation factor VII F7 P08833 Insulin-like growth factor-binding protein 1 IGFBP1 P08887 Interleukin-6 receptor subunit alpha IL6R P08949 Neuromedin-B-32 NMB P08F94 Fibrocystin PKHD1 P09038 Fibroblast growth factor 2 FGF2 P09228 Cystatin-SA CST2 P09237 Matrilysin MMP7 P09238 Stromelysin-2 MMP10 P09341 Growth-regulated alpha protein CXCL1 P09382 Galectin-1 LGALS1 P09466 Glycodelin PAEP P09486 SPARC SPARC P09529 Inhibin beta B chain INHBB P09544 Protein Wnt-2 WNT2 P09603 Processed macrophage colony-stimulating factor 1 CSF1 P09681 Gastric inhibitory polypeptide GIP P09683 Secretin SCT P09919 Granulocyte colony-stimulating factor CSF3 P0C091 FRAS1-related extracellular matrix protein 3 FREM3 P0C0L4 C4d-A C4A P0C0L5 Complement C4-B alpha chain C4B P0C0P6 Neuropeptide S NPS P0C7L1 Serine protease inhibitor Kazal-type 8 SPINK8 P0C862 Complement C1q and tumor necrosis factor- C1QTNF9 related protein 9A P0C8F1 Prostate and testis expressed protein 4 PATE4 P0CG01 Gastrokine-3 GKN3P P0CG36 Cryptic family protein 1B CFC1B P0CG37 Cryptic protein CFC1 P0CJ68 Humanin-like protein 1 MTRNR2L1 P0CJ69 Humanin-like protein 2 MTRNR2L2 P0CJ70 Humanin-like protein 3 MTRNR2L3 P0CJ71 Humanin-like protein 4 MTRNR2L4 P0CJ72 Humanin-like protein 5 MTRNR2L5 P0CJ73 Humanin-like protein 6 MTRNR2L6 P0CJ74 Humanin-like protein 7 MTRNR2L7 P0CJ75 Humanin-like protein 8 MTRNR2L8 P0CJ76 Humanin-like protein 9 MTRNR2L9 P0CJ77 Humanin-like protein 10 MTRNR2L10 P0DJD7 Pepsin A-4 PGA4 P0DJD8 Pepsin A-3 PGA3 P0DJD9 Pepsin A-5 PGA5 P0DJI8 Amyloid protein A SAA1 P0DJI9 Serum amyloid A-2 protein SAA2 P10082 Peptide YY(3-36) PYY P10092 Calcitonin gene-related peptide 2 CALCB P10124 Serglycin SRGN P10145 MDNCF-a IL8 P10147 MIP-1-alpha(4-69) CCL3 P10163 Peptide P-D PRB4 P10451 Osteopontin SPP1 P10599 Thioredoxin TXN P10600 Transforming growth factor beta-3 TGFB3 P10643 Complement component C7 C7 P10645 Vasostatin-2 CHGA P10646 Tissue factor pathway inhibitor TFPI P10720 Platelet factor 4 variant(4-74) PF4V1 P10745 Retinol-binding protein 3 RBP3 P10767 Fibroblast growth factor 6 FGF6 P10909 Clusterin alpha chain CLU P10912 Growth hormone receptor GHR P10915 Hyaluronan and proteoglycan link protein 1 HAPLN1 P10966 T-cell surface glycoprotein CD8 beta chain CD8B P10997 Islet amyloid polypeptide IAPP P11047 Laminin subunit gamma-1 LAMC1 P11150 Hepatic triacylglycerol lipase LIPC P11226 Mannose-binding protein C MBL2 P11464 Pregnancy-specific beta-1-glycoprotein 1 PSG1 P11465 Pregnancy-specific beta-1-glycoprotein 2 PSG2 P11487 Fibroblast growth factor 3 FGF3 P11597 Cholesteryl ester transfer protein CETP P11684 Uteroglobin SCGB1A1 P11686 Pulmonary surfactant-associated protein C SFTPC P12034 Fibroblast growth factor 5 FGF5 P12107 Collagen alpha-1(XI) chain COL11A1 P12109 Collagen alpha-1(VI) chain COL6A1 P12110 Collagen alpha-2(VI) chain COL6A2 P12111 Collagen alpha-3(VI) chain COL6A3 P12259 Coagulation factor V F5 P12272 PTHrP[1-36] PTHLH P12273 Prolactin-inducible protein PIP P12544 Granzyme A GZMA P12643 Bone morphogenetic protein 2 BMP2 P12644 Bone morphogenetic protein 4 BMP4 P12645 Bone morphogenetic protein 3 BMP3 P12724 Eosinophil cationic protein RNASE3 P12821 Angiotensin-converting enzyme, soluble form ACE P12838 Neutrophil defensin 4 DEFA4 P12872 Motilin MLN P13232 Interleukin-7 IL7 P13236 C-C motif chemokine 4 CCL4 P13284 Gamma-interferon-inducible lysosomal thiol IFI30 reductase P13500 C-C motif chemokine 2 CCL2 P13501 C-C motif chemokine 5 CCL5 P13521 Secretogranin-2 SCG2 P13591 Neural cell adhesion molecule 1 NCAM1 P13611 Versican core protein VCAN P13671 Complement component C6 C6 P13688 Carcinoembryonic antigen-related cell adhesion CEACAM1 molecule 1 P13725 Oncostatin-M OSM P13726 Tissue factor F3 P13727 Eosinophil granule major basic protein PRG2 P13942 Collagen alpha-2(XI) chain COL11A2 P13987 CD59 glycoprotein CD59 P14138 Endothelin-3 EDN3 P14174 Macrophage migration inhibitory factor MIF P14207 Folate receptor beta FOLR2 P14222 Perforin-1 PRF1 P14543 Nidogen-1 NID1 P14555 Phospholipase A2, membrane associated PLA2G2A P14625 Endoplasmin HSP90B1 P14735 Insulin-degrading enzyme IDE P14778 Interleukin-1 receptor type 1, soluble form IL1R1 P14780 82 kDa matrix metalloproteinase-9 MMP9 P15018 Leukemia inhibitory factor LIF P15085 Carboxypeptidase A1 CPA1 P15086 Carboxypeptidase B CPB1 P15151 Poliovirus receptor PVR P15169 Carboxypeptidase N catalytic chain CPN1 P15248 Interleukin-9 IL9 P15291 N-acetyllactosamine synthase B4GALT1 P15309 PAPG9 ACPP P15328 Folate receptor alpha FOLR1 P15374 Ubiquitin carboxyl-terminal hydrolase isozyme L3 UCHL3 P15502 Elastin ELN P15509 Granulocyte-macrophage colony-stimulating CSF2RA factor receptor subunit alpha P15515 Histatin-1 HTN1 P15516 His3-(31-51)-peptide HTN3 P15692 Vascular endothelial growth factor A VEGFA P15814 Immunoglobulin lambda-like polypeptide 1 IGLL1 P15907 Beta-galactoside alpha-2,6-sialyltransferase 1 ST6GAL1 P15941 Mucin-1 subunit beta MUC1 P16035 Metalloproteinase inhibitor 2 TIMP2 P16112 Aggrecan core protein 2 ACAN P16233 Pancreatic triacylglycerol lipase PNLIP P16442 Histo-blood group ABO system transferase ABO P16471 Prolactin receptor PRLR P16562 Cysteine-rich secretory protein 2 CRISP2 P16619 C-C motif chemokine 3-like 1 CCL3L1 P16860 BNP(3-29) NPPB P16870 Carboxypeptidase E CPE P16871 Interleukin-7 receptor subunit alpha IL7R P17213 Bactericidal permeability-increasing protein BPI P17538 Chymotrypsinogen B CTRB1 P17931 Galectin-3 LGALS3 P17936 Insulin-like growth factor-binding protein 3 IGFBP3 P17948 Vascular endothelial growth factor receptor 1 FLT1 P18065 Insulin-like growth factor-binding protein 2 IGFBP2 P18075 Bone morphogenetic protein 7 BMP7 P18428 Lipopolysaccharide-binding protein LBP P18509 PACAP-related peptide ADCYAP1 P18510 Interleukin-1 receptor antagonist protein IL1RN P18827 Syndecan-1 SDC1 P19021 Peptidylglycine alpha-hydroxylating PAM monooxygenase P19235 Erythropoietin receptor EPOR P19438 Tumor necrosis factor-binding protein 1 TNFRSF1A P19652 Alpha-1-acid glycoprotein 2 ORM2 P19801 Amiloride-sensitive amine oxidase [copper- ABP1 containing] P19823 Inter-alpha-trypsin inhibitor heavy chain H2 ITIH2 P19827 Inter-alpha-trypsin inhibitor heavy chain H1 ITIH1 P19835 Bile salt-activated lipase CEL P19875 C-X-C motif chemokine 2 CXCL2 P19876 C-X-C motif chemokine 3 CXCL3 P19883 Follistatin FST P19957 Elafin PI3 P19961 Alpha-amylase 2B AMY2B P20061 Transcobalamin-1 TCN1 P20062 Transcobalamin-2 TCN2 P20142 Gastricsin PGC P20155 Serine protease inhibitor Kazal-type 2 SPINK2 P20231 Tryptase beta-2 TPSB2 P20333 Tumor necrosis factor receptor superfamily TNFRSF1B member 1B P20366 Substance P TAC1 P20382 Melanin-concentrating hormone PMCH P20396 Thyroliberin TRH P20742 Pregnancy zone protein PZP P20774 Mimecan OGN P20783 Neurotrophin-3 NTF3 P20800 Endothelin-2 EDN2 P20809 Interleukin-11 IL11 P20827 Ephrin-A1 EFNA1 P20849 Collagen alpha-1(IX) chain COL9A1 P20851 C4b-binding protein beta chain C4BPB P20908 Collagen alpha-1(V) chain COL5A1 P21128 Poly(U)-specific endoribonuclease ENDOU P21246 Pleiotrophin PTN P21583 Kit ligand KITLG P21741 Midkine MDK P21754 Zona pellucida sperm-binding protein 3 ZP3 P21781 Fibroblast growth factor 7 FGF7 P21802 Fibroblast growth factor receptor 2 FGFR2 P21810 Biglycan BGN P21815 Bone sialoprotein 2 IBSP P21860 Receptor tyrosine-protein kinase erbB-3 ERBB3 P21941 Cartilage matrix protein MATN1 P22003 Bone morphogenetic protein 5 BMP5 P22004 Bone morphogenetic protein 6 BMP6 P22079 Lactoperoxidase LPO P22105 Tenascin-X TNXB P22301 Interleukin-10 IL10 P22303 Acetylcholinesterase ACHE P22352 Glutathione peroxidase 3 GPX3 P22362 C-C motif chemokine 1 CCL1 P22455 Fibroblast growth factor receptor 4 FGFR4 P22466 Galanin message-associated peptide GAL P22692 Insulin-like growth factor-binding protein 4 IGFBP4 P22749 Granulysin GNLY P22792 Carboxypeptidase N subunit 2 CPN2 P22891 Vitamin K-dependent protein Z PROZ P22894 Neutrophil collagenase MMP8 P23142 Fibulin-1 FBLN1 P23280 Carbonic anhydrase 6 CA6 P23352 Anosmin-1 KAL1 P23435 Cerebellin-1 CBLN1 P23560 Brain-derived neurotrophic factor BDNF P23582 C-type natriuretic peptide NPPC P23946 Chymase CMA1 P24043 Laminin subunit alpha-2 LAMA2 P24071 Immunoglobulin alpha Fc receptor FCAR P24347 Stromelysin-3 MMP11 P24387 Corticotropin-releasing factor-binding protein CRHBP P24592 Insulin-like growth factor-binding protein 6 IGFBP6 P24593 Insulin-like growth factor-binding protein 5 IGFBP5 P24821 Tenascin TNC P24855 Deoxyribonuclease-1 DNASE1 P25067 Collagen alpha-2(VIII) chain COL8A2 P25311 Zinc-alpha-2-glycoprotein AZGP1 P25391 Laminin subunit alpha-1 LAMA1 P25445 Tumor necrosis factor receptor superfamily FAS member 6 P25940 Collagen alpha-3(V) chain COL5A3 P25942 Tumor necrosis factor receptor superfamily CD40 member 5 P26022 Pentraxin-related protein PTX3 PTX3 P26927 Hepatocyte growth factor-like protein beta chain MST1 P27169 Serum paraoxonase/arylesterase 1 PON1 P27352 Gastric intrinsic factor GIF P27487 Dipeptidyl peptidase 4 membrane form DPP4 P27539 Embryonic growth/differentiation factor 1 GDF1 P27658 Vastatin COL8A1 P27797 Calreticulin CALR P27918 Properdin CFP P28039 Acyloxyacyl hydrolase AOAH P28300 Protein-lysine 6-oxidase LOX P28325 Cystatin-D CST5 P28799 Granulin-1 GRN P29122 Proprotein convertase subtilisin/kexin type 6 PCSK6 P29279 Connective tissue growth factor CTGF P29320 Ephrin type-A receptor 3 EPHA3 P29400 Collagen alpha-5(IV) chain COL4A5 P29459 Interleukin-12 subunit alpha IL12A P29460 Interleukin-12 subunit beta IL12B P29508 Serpin B3 SERPINB3 P29622 Kallistatin SERPINA4 P29965 CD40 ligand, soluble form CD40LG P30990 Neurotensin/neuromedin N NTS P31025 Lipocalin-1 LCN1 P31151 Protein S100-A7 S100A7 P31371 Fibroblast growth factor 9 FGF9 P31431 Syndecan-4 SDC4 P31947 14-3-3 protein sigma SFN P32455 Interferon-induced guanylate-binding protein 1 GBP1 P32881 Interferon alpha-8 IFNA8 P34096 Ribonuclease 4 RNASE4 P34130 Neurotrophin-4 NTF4 P34820 Bone morphogenetic protein 8B BMP8B P35030 Trypsin-3 PRSS3 P35052 Secreted glypican-1 GPC1 P35070 Betacellulin BTC P35225 Interleukin-13 IL13 P35247 Pulmonary surfactant-associated protein D SFTPD P35318 ADM ADM P35542 Serum amyloid A-4 protein SAA4 P35555 Fibrillin-1 FBN1 P35556 Fibrillin-2 FBN2 P35625 Metalloproteinase inhibitor 3 TIMP3 P35858 Insulin-like growth factor-binding protein complex IGFALS acid labile subunit P35916 Vascular endothelial growth factor receptor 3 FLT4 P35968 Vascular endothelial growth factor receptor 2 KDR P36222 Chitinase-3-like protein 1 CHI3L1 P36952 Serpin B5 SERPINB5 P36955 Pigment epithelium-derived factor SERPINF1 P36980 Complement factor H-related protein 2 CFHR2 P39059 Collagen alpha-1(XV) chain COL15A1 P39060 Collagen alpha-1(XVIII) chain COL18A1 P39877 Calcium-dependent phospholipase A2 PLA2G5 P39900 Macrophage metalloelastase MMP12 P39905 Glial cell line-derived neurotrophic factor GDNF P40225 Thrombopoietin THPO P40967 M-alpha PMEL P41159 Leptin LEP P41221 Protein Wnt-5a WNT5A P41222 Prostaglandin-H2 D-isomerase PTGDS P41271 Neuroblastoma suppressor of tumorigenicity 1 NBL1 P41439 Folate receptor gamma FOLR3 P42127 Agouti-signaling protein ASIP P42702 Leukemia inhibitory factor receptor LIFR P42830 ENA-78(9-78) CXCL5 P43026 Growth/differentiation factor 5 GDF5 P43251 Biotinidase BTD P43652 Afamin AFM P45452 Collagenase 3 MMP13 P47710 Casoxin-D CSN1S1 P47929 Galectin-7 LGALS7B P47972 Neuronal pentraxin-2 NPTX2 P47989 Xanthine oxidase XDH P47992 Lymphotactin XCL1 P48023 Tumor necrosis factor ligand superfamily member FASLG 6, membrane form P48052 Carboxypeptidase A2 CPA2 P48061 Stromal cell-derived factor 1 CXCL12 P48304 Lithostathine-1-beta REG1B P48307 Tissue factor pathway inhibitor 2 TFPI2 P48357 Leptin receptor LEPR P48594 Serpin B4 SERPINB4 P48645 Neuromedin-U-25 NMU P48740 Mannan-binding lectin serine protease 1 MASP1 P48745 Protein NOV homolog NOV P48960 CD97 antigen subunit beta CD97 P49223 Kunitz-type protease inhibitor 3 SPINT3 P49747 Cartilage oligomeric matrix protein COMP P49763 Placenta growth factor PGF P49765 Vascular endothelial growth factor B VEGFB P49767 Vascular endothelial growth factor C VEGFC P49771 Fms-related tyrosine kinase 3 ligand FLT3LG P49862 Kallikrein-7 KLK7 P49863 Granzyme K GZMK P49908 Selenoprotein P SEPP1 P49913 Antibacterial protein FALL-39 CAMP P50607 Tubby protein homolog TUB P51124 Granzyme M GZMM P51512 Matrix metalloproteinase-16 MMP16 P51654 Glypican-3 GPC3 P51671 Eotaxin CCL11 P51884 Lumican LUM P51888 Prolargin PRELP P52798 Ephrin-A4 EFNA4 P52823 Stanniocalcin-1 STC1 P53420 Collagen alpha-4(IV) chain COL4A4 P53621 Coatomer subunit alpha COPA P54108 Cysteine-rich secretory protein 3 CRISP3 P54315 Pancreatic lipase-related protein 1 PNLIPRP1 P54317 Pancreatic lipase-related protein 2 PNLIPRP2 P54793 Arylsulfatase F ARSF P55000 Secreted Ly-6/uPAR-related protein 1 SLURP 1 P55001 Microfibrillar-associated protein 2 MFAP2 P55056 Apolipoprotein C-IV APOC4 P55058 Phospholipid transfer protein PLTP P55075 Fibroblast growth factor 8 FGF8 P55081 Microfibrillar-associated protein 1 MFAP1 P55083 Microfibril-associated glycoprotein 4 MFAP4 P55107 Bone morphogenetic protein 3B GDF10 P55145 Mesencephalic astrocyte-derived neurotrophic MANF factor P55259 Pancreatic secretory granule membrane major GP2 glycoprotein GP2 P55268 Laminin subunit beta-2 LAMB2 P55773 CCL23(30-99) CCL23 P55774 C-C motif chemokine 18 CCL18 P55789 FAD-linked sulfhydryl oxidase ALR GFER P56703 Proto-oncogene Wnt-3 WNT3 P56704 Protein Wnt-3a WNT3A P56705 Protein Wnt-4 WNT4 P56706 Protein Wnt-7b WNT7B P56730 Neurotrypsin PRSS12 P56851 Epididymal secretory protein E3-beta EDDM3B P56975 Neuregulin-3 NRG3 P58062 Serine protease inhibitor Kazal-type 7 SPINK7 P58215 Lysyl oxidase homolog 3 LOXL3 P58294 Prokineticin-1 PROK1 P58335 Anthrax toxin receptor 2 ANTXR2 P58397 A disintegrin and metalloproteinase with ADAMTS12 thrombospondin motifs 12 P58417 Neurexophilin-1 NXPH1 P58499 Protein FAM3B FAM3B P59510 A disintegrin and metalloproteinase with ADAMTS20 thrombospondin motifs 20 P59665 Neutrophil defensin 1 DEFA1B P59666 Neutrophil defensin 3 DEFA3 P59796 Glutathione peroxidase 6 GPX6 P59826 BPI fold-containing family B member 3 BPIFB3 P59827 BPI fold-containing family B member 4 BPIFB4 P59861 Beta-defensin 131 DEFB131 P60022 Beta-defensin 1 DEFB1 P60153 Inactive ribonuclease-like protein 9 RNASE9 P60827 Complement C1q tumor necrosis factor-related C1QTNF8 protein 8 P60852 Zona pellucida sperm-binding protein 1 ZP1 P60985 Keratinocyte differentiation-associated protein KRTDAP P61109 Kidney androgen-regulated protein KAP P61278 Somatostatin-14 SST P61366 Osteocrin OSTN P61626 Lysozyme C LYZ P61769 Beta-2-microglobulin B2M P61812 Transforming growth factor beta-2 TGFB2 P61916 Epididymal secretory protein E1 NPC2 P62502 Epididymal-specific lipocalin-6 LCN6 P62937 Peptidyl-prolyl cis-trans isomerase A PPIA P67809 Nuclease-sensitive element-binding protein 1 YBX1 P67812 Signal peptidase complex catalytic subunit SEC11A SEC11A P78310 Coxsackievirus and adenovirus receptor CXADR P78333 Secreted glypican-5 GPC5 P78380 Oxidized low-density lipoprotein receptor 1 OLR1 P78423 Processed fractalkine CX3CL1 P78509 Reelin RELN P78556 CCL20(2-70) CCL20 P80075 MCP-2(6-76) CCL8 P80098 C-C motif chemokine 7 CCL7 P80108 Phosphatidylinositol-glycan-specific GPLD1 phospholipase D P80162 C-X-C motif chemokine 6 CXCL6 P80188 Neutrophil gelatinase-associated lipocalin LCN2 P80303 Nucleobindin-2 NUCB2 P80511 Calcitermin S100A12 P81172 Hepcidin-25 HAMP P81277 Prolactin-releasing peptide PRLH P81534 Beta-defensin 103 DEFB103A P81605 Dermcidin DCD P82279 Protein crumbs homolog 1 CRB1 P82987 ADAMTS-like protein 3 ADAMTSL3 P83105 Serine protease HTRA4 HTRA4 P83110 Serine protease HTRA3 HTRA3 P83859 Orexigenic neuropeptide QRFP QRFP P98088 Mucin-5AC MUC5AC P98095 Fibulin-2 FBLN2 P98160 Basement membrane-specific heparan sulfate HSPG2 proteoglycan core protein P98173 Protein FAM3A FAM3A Q00604 Norrin NDP Q00796 Sorbitol dehydrogenase SORD Q00887 Pregnancy-specific beta-1-glycoprotein 9 PSG9 Q00888 Pregnancy-specific beta-1-glycoprotein 4 PSG4 Q00889 Pregnancy-specific beta-1-glycoprotein 6 PSG6 Q01523 HD5(56-94) DEFA5 Q01524 Defensin-6 DEFA6 Q01955 Collagen alpha-3(IV) chain COL4A3 Q02297 Pro-neuregulin-1, membrane-bound isoform NRG1 Q02325 Plasminogen-like protein B PLGLB1 Q02383 Semenogelin-2 SEMG2 Q02388 Collagen alpha-1(VII) chain COL7A1 Q02505 Mucin-3A MUC3A Q02509 Otoconin-90 OC90 Q02747 Guanylin GUCA2A Q02763 Angiopoietin-1 receptor TEK Q02817 Mucin-2 MUC2 Q02985 Complement factor H-related protein 3 CFHR3 Q03167 Transforming growth factor beta receptor type 3 TGFBR3 Q03403 Trefoil factor 2 TFF2 Q03405 Urokinase plasminogen activator surface receptor PLAUR Q03591 Complement factor H-related protein 1 CFHR1 Q03692 Collagen alpha-1(X) chain COL10A1 Q04118 Basic salivary proline-rich protein 3 PRB3 Q04756 Hepatocyte growth factor activator short chain HGFAC Q04900 Sialomucin core protein 24 CD164 Q05315 Eosinophil lysophospholipase CLC Q05707 Collagen alpha-1 (XIV) chain COL14A1 Q05996 Processed zona pellucida sperm-binding protein 2 ZP2 Q06033 Inter-alpha-trypsin inhibitor heavy chain H3 ITIH3 Q06141 Regenerating islet-derived protein 3-alpha REG3A Q06828 Fibromodulin FMOD Q07092 Collagen alpha-1(XVI) chain COL16A1 Q07325 C-X-C motif chemokine 9 CXCL9 Q07507 Dermatopontin DPT Q075Z2 Binder of sperm protein homolog 1 BSPH1 Q07654 Trefoil factor 3 TFF3 Q07699 Sodium channel subunit beta-1 SCN1B Q08345 Epithelial discoidin domain-containing receptor 1 DDR1 Q08380 Galectin-3-binding protein LGALS3BP Q08397 Lysyl oxidase homolog 1 LOXL1 Q08431 Lactadherin MFGE8 Q08629 Testican-1 SPOCK1 Q08648 Sperm-associated antigen 11B SPAG11B Q08830 Fibrinogen-like protein 1 FGL1 Q10471 Polypeptide N-acetylgalactosaminyltransferase 2 GALNT2 Q10472 Polypeptide N-acetylgalactosaminyltransferase 1 GALNT1 Q11201 CMP-N-acetylneuraminate-beta-galactosamide- ST3GAL1 alpha-2,3-sialyltransferase 1 Q11203 CMP-N-acetylneuraminate-beta-1,4-galactoside ST3GAL3 alpha-2,3-sialyltransferase Q11206 CMP-N-acetylneuraminate-beta-galactosamide- ST3GAL4 alpha-2,3-sialyltransferase 4 Q12794 Hyaluronidase-1 HYAL1 Q12805 EGF-containing fibulin-like extracellular matrix EFEMP1 protein 1 Q12836 Zona pellucida sperm-binding protein 4 ZP4 Q12841 Follistatin-related protein 1 FSTL1 Q12904 Aminoacyl tRNA synthase complex-interacting AIMP1 multifunctional protein 1 Q13018 Soluble secretory phospholipase A2 receptor PLA2R1 Q13072 B melanoma antigen 1 BAGE Q13093 Platelet-activating factor acetylhydrolase PLA2G7 Q13103 Secreted phosphoprotein 24 SPP2 Q13162 Peroxiredoxin-4 PRDX4 Q13201 Platelet glycoprotein Ia* MMRN1 Q13214 Semaphorin-3B SEMA3B Q13219 Pappalysin-1 PAPPA Q13231 Chitotriosidase-1 CHIT1 Q13253 Noggin NOG Q13261 Interleukin-15 receptor subunit alpha IL15RA Q13275 Semaphorin-3F SEMA3F Q13291 Signaling lymphocytic activation molecule SLAMF1 Q13316 Dentin matrix acidic phosphoprotein 1 DMP1 Q13361 Microfibrillar-associated protein 5 MFAP5 Q13410 Butyrophilin subfamily 1 member A1 BTN1A1 Q13421 Mesothelin, cleaved form MSLN Q13429 Insulin-like growth factor I IGF-I Q13443 Disintegrin and metalloproteinase domain- ADAM9 containing protein 9 Q13519 Neuropeptide 1 PNOC Q13751 Laminin subunit beta-3 LAMB3 Q13753 Laminin subunit gamma-2 LAMC2 Q13790 Apolipoprotein F APOF Q13822 Ectonucleotide ENPP2 pyrophosphatase/phosphodiesterase family member 2 Q14031 Collagen alpha-6(IV) chain COL4A6 Q14050 Collagen alpha-3(IX) chain COL9A3 Q14055 Collagen alpha-2(IX) chain COL9A2 Q14112 Nidogen-2 NID2 Q14114 Low-density lipoprotein receptor-related protein 8 LRP8 Q14118 Dystroglycan DAG1 Q14314 Fibroleukin FGL2 Q14393 Growth arrest-specific protein 6 GAS6 Q14406 Chorionic somatomammotropin hormone-like 1 CSHL1 Q14507 Epididymal secretory protein E3-alpha EDDM3A Q14508 WAP four-disulfide core domain protein 2 WFDC2 Q14512 Fibroblast growth factor-binding protein 1 FGFBP1 Q14515 SPARC-like protein 1 SPARCL1 Q14520 Hyaluronan-binding protein 2 27 kDa light chain HABP2 Q14563 Semaphorin-3A SEMA3A Q14623 Indian hedgehog protein IHH Q14624 Inter-alpha-trypsin inhibitor heavy chain H4 ITIH4 Q14667 UPF0378 protein KIAA0100 KIAA0100 Q14703 Membrane-bound transcription factor site-1 MBTPS1 protease Q14766 Latent-transforming growth factor beta-binding LTBP1 protein 1 Q14767 Latent-transforming growth factor beta-binding LTBP2 protein 2 Q14773 Intercellular adhesion molecule 4 ICAM4 Q14993 Collagen alpha-1(XIX) chain COL19A1 Q14CN2 Calcium-activated chloride channel regulator 4, CLCA4 110 kDa form Q15046 Lysine--tRNA ligase KARS Q15063 Periostin POSTN Q15109 Advanced glycosylation end product-specific AGER receptor Q15113 Procollagen C-endopeptidase enhancer 1 PCOLCE Q15166 Serum paraoxonase/lactonase 3 PON3 Q15195 Plasminogen-like protein A PLGLA Q15198 Platelet-derived growth factor receptor-like protein PDGFRL Q15223 Poliovirus receptor-related protein 1 PVRL1 Q15238 Pregnancy-specific beta-1-glycoprotein 5 PSG5 Q15363 Transmembrane emp24 domain-containing protein 2 TMED2 Q15375 Ephrin type-A receptor 7 EPHA7 Q15389 Angiopoietin-1 ANGPT1 Q15465 Sonic hedgehog protein SHH Q15485 Ficolin-2 FCN2 Q15517 Corneodesmosin CDSN Q15582 Transforming growth factor-beta-induced protein TGFBI ig-h3 Q15661 Tryptase alpha/beta-1 TPSAB1 Q15726 Metastin KISS1 Q15782 Chitinase-3-like protein 2 CHI3L2 Q15828 Cystatin-M CST6 Q15846 Clusterin-like protein 1 CLUL1 Q15848 Adiponectin ADIPOQ Q16206 Protein disulfide-thiol oxidoreductase ENOX2 Q16270 Insulin-like growth factor-binding protein 7 IGFBP7 Q16363 Laminin subunit alpha-4 LAMA4 Q16378 Proline-rich protein 4 PRR4 Q16557 Pregnancy-specific beta-1-glycoprotein 3 PSG3 Q16568 CART(42-89) CARTPT Q16610 Extracellular matrix protein 1 ECM1 Q16619 Cardiotrophin-1 CTF1 Q16623 Syntaxin-1A STX1A Q16627 HCC-1(9-74) CCL14 Q16651 Prostasin light chain PRSS8 Q16661 Guanylate cyclase C-activating peptide 2 GUCA2B Q16663 CCL15(29-92) CCL15 Q16674 Melanoma-derived growth regulatory protein MIA Q16769 Glutaminyl-peptide cyclotransferase QPCT Q16787 Laminin subunit alpha-3 LAMA3 Q16842 CMP-N-acetylneuraminate-beta-galactosamide- ST3GAL2 alpha-2,3-sialyltransferase 2 Q17RR3 Pancreatic lipase-related protein 3 PNLIPRP3 Q17RW2 Collagen alpha-1(XXIV) chain COL24A1 Q17RY6 Lymphocyte antigen 6K LY6K Q1L6U9 Prostate-associated microseminoprotein MSMP Q1W4C9 Serine protease inhibitor Kazal-type 13 SPINK13 Q1ZYL8 Izumo sperm-egg fusion protein 4 IZUMO4 Q29960 HLA class I histocompatibility antigen, Cw-16 HLA-C alpha chain Q2I0M5 R-spondin-4 RSPO4 Q2L4Q9 Serine protease 53 PRSS53 Q2MKA7 R-spondin-1 RSPO1 Q2MV58 Tectonic-1 TCTN1 Q2TAL6 Brorin VWC2 Q2UY09 Collagen alpha-1(XXVIII) chain COL28A1 Q2VPA4 Complement component receptor 1-like protein CR1L Q2WEN9 Carcinoembryonic antigen-related cell adhesion CEACAM16 molecule 16 Q30KP8 Beta-defensin 136 DEFB136 Q30KP9 Beta-defensin 135 DEFB135 Q30KQ1 Beta-defensin 133 DEFB133 Q30KQ2 Beta-defensin 130 DEFB130 Q30KQ4 Beta-defensin 116 DEFB116 Q30KQ5 Beta-defensin 115 DEFB115 Q30KQ6 Beta-defensin 114 DEFB114 Q30KQ7 Beta-defensin 113 DEFB113 Q30KQ8 Beta-defensin 112 DEFB112 Q30KQ9 Beta-defensin 110 DEFB110 Q30KR1 Beta-defensin 109 DEFB109P1 Q32P28 Prolyl 3-hydroxylase 1 LEPRE1 Q3B7J2 Glucose-fructose oxidoreductase domain- GFOD2 containing protein 2 Q3SY79 Protein Wnt WNT3A Q3T906 N-acetylglucosamine-1-phosphotransferase GNPTAB subunits alpha/beta Q495T6 Membrane metallo-endopeptidase-like 1 MMEL1 Q49AH0 Cerebral dopamine neurotrophic factor CDNF Q4G0G5 Secretoglobin family 2B member 2 SCGB2B2 Q4G0M1 Protein FAM132B FAM132B Q4LDE5 Sushi, von Willebrand factor type A, EGF and SVEP1 pentraxin domain-containing protein 1 Q4QY38 Beta-defensin 134 DEFB134 Q4VAJ4 Protein Wnt WNT10B Q4W5P6 Protein TMEM155 TMEM155 Q4ZHG4 Fibronectin type III domain-containing protein 1 FNDC1 Q53H76 Phospholipase A1 member A PLA1A Q53RD9 Fibulin-7 FBLN7 Q53S33 BolA-like protein 3 BOLA3 Q5BLP8 Neuropeptide-like protein C4orf48 C4orf48 Q5DT21 Serine protease inhibitor Kazal-type 9 SPINK9 Q5EBL8 PDZ domain-containing protein 11 PDZD11 Q5FYB0 Arylsulfatase J ARSJ Q5FYB1 Arylsulfatase I ARSI Q5GAN3 Ribonuclease-like protein 13 RNASE13 Q5GAN4 Ribonuclease-like protein 12 RNASE12 Q5GAN6 Ribonuclease-like protein 10 RNASE10 Q5GFL6 von Willebrand factor A domain-containing VWA2 protein 2 Q5H8A3 Neuromedin-S NMS Q5H8C1 FRAS1-related extracellular matrix protein 1 FREM1 Q5IJ48 Protein crumbs homolog 2 CRB2 Q5J5C9 Beta-defensin 121 DEFB121 Q5JS37 NHL repeat-containing protein 3 NHLRC3 Q5JTB6 Placenta-specific protein 9 PLAC9 Q5JU69 Torsin-2A TOR2A Q5JXM2 Methyltransferase-like protein 24 METTL24 Q5JZY3 Ephrin type-A receptor 10 EPHA10 Q5K4E3 Polyserase-2 PRSS36 Q5SRR4 Lymphocyte antigen 6 complex locus protein G5c LY6G5C Q5T1H1 Protein eyes shut homolog EYS Q5T4F7 Secreted frizzled-related protein 5 SFRP5 Q5T4W7 Artemin ARTN Q5T7M4 Protein FAM132A FAM132A Q5TEH8 Protein Wnt WNT2B Q5TIE3 von Willebrand factor A domain-containing VWA5B1 protein 5B1 Q5UCC4 ER membrane protein complex subunit 10 EMC10 Q5VST6 Abhydrolase domain-containing protein FAM108B1 FAM108B1 Q5VTL7 Fibronectin type III domain-containing protein 7 FNDC7 Q5VUM1 UPF0369 protein C6orf57 C6orf57 Q5VV43 Dyslexia-associated protein KIAA0319 KIAA0319 Q5VWW1 Complement C1q-like protein 3 C1QL3 Q5VXI9 Lipase member N LIPN Q5VXJ0 Lipase member K LIPK Q5VXM1 CUB domain-containing protein 2 CDCP2 Q5VYX0 Renalase RNLS Q5VYY2 Lipase member M LIPM Q5W186 Cystatin-9 CST9 Q5W5W9 Regulated endocrine-specific protein 18 RESP18 Q5XG92 Carboxylesterase 4A CES4A Q63HQ2 Pikachurin EGFLAM Q641Q3 Meteorin-like protein METRNL Q66K79 Carboxypeptidase Z CPZ Q685J3 Mucin-17 MUC17 Q68BL7 Olfactomedin-like protein 2A OLFML2A Q68BL8 Olfactomedin-like protein 2B OLFML2B Q68DV7 E3 ubiquitin-protein ligase RNF43 RNF43 Q6B9Z1 Insulin growth factor-like family member 4 IGFL4 Q6BAA4 Fc receptor-like B FCRLB Q6E0U4 Dermokine DMKN Q6EMK4 Vasorin VASN Q6FHJ7 Secreted frizzled-related protein 4 SFRP4 Q6GPI1 Chymotrypsin B2 chain B CTRB2 Q6GTS8 Probable Carboxypeptidase PM20D1 PM20D1 Q6H9L7 Isthmin-2 ISM2 Q6IE36 Ovostatin homolog 2 OVOS2 Q6IE37 Ovostatin homolog 1 OVOS1 Q6IE38 Serine protease inhibitor Kazal-type 14 SPINK14 Q6ISS4 Leukocyte-associated immunoglobulin-like LAIR2 receptor 2 Q6JVE5 Epididymal-specific lipocalin-12 LCN12 Q6JVE6 Epididymal-specific lipocalin-10 LCN10 Q6JVE9 Epididymal-specific lipocalin-8 LCN8 Q6KF10 Growth/differentiation factor 6 GDF6 Q6MZW2 Follistatin-related protein 4 FSTL4 Q6NSX1 Coiled-coil domain-containing protein 70 CCDC70 Q6NT32 Carboxylesterase 5A CES5A Q6NT52 Choriogonadotropin subunit beta variant 2 CGB2 Q6NUI6 Chondroactherin-like protein CHADL Q6NUJ1 Saposin A-like PSAPL1 Q6P093 Arylacetamide deacetylase-like 2 AADACL2 Q6P4A8 Phospholipase B-like 1 PLBD1 Q6P5S2 UPF0762 protein C6orf58 C6orf58 Q6P988 Protein notum homolog NOTUM Q6PCB0 von Willebrand factor A domain-containing VWA1 protein 1 Q6PDA7 Sperm-associated antigen 11A SPAG11A Q6PEW0 Inactive serine protease 54 PRSS54 Q6PEZ8 Podocan-like protein 1 PODNL1 Q6PKH6 Dehydrogenase/reductase SDR family member 4- DHRS4L2 like 2 Q6Q788 Apolipoprotein A-V APOA5 Q6SPF0 Atherin SAMD1 Q6UDR6 Kunitz-type protease inhibitor 4 SPINT4 Q6URK8 Testis, prostate and placenta-expressed protein TEPP Q6UW01 Cerebellin-3 CBLN3 Q6UW10 Surfactant-associated protein 2 SFTA2 Q6UW15 Regenerating islet-derived protein 3-gamma REG3G Q6UW32 Insulin growth factor-like family member 1 IGFL1 Q6UW78 UPF0723 protein C11orf83 C11orf83 Q6UW88 Epigen EPGN Q6UWE3 Colipase-like protein 2 CLPSL2 Q6UWF7 NXPE family member 4 NXPE4 Q6UWF9 Protein FAM180A FAM180A Q6UWM5 GLIPR1-like protein 1 GLIPR1L1 Q6UWN8 Serine protease inhibitor Kazal-type 6 SPINK6 Q6UWP2 Dehydrogenase/reductase SDR family member 11 DHRS11 Q6UWP8 Suprabasin SBSN Q6UWQ5 Lysozyme-like protein 1 LYZL1 Q6UWQ7 Insulin growth factor-like family member 2 IGFL2 Q6UWR7 Ectonucleotide ENPP6 pyrophosphatase/phosphodiesterase family member 6 soluble form Q6UWT2 Adropin ENHO Q6UWU2 Beta-galactosidase-1-like protein GLB1L Q6UWW0 Lipocalin-15 LCN15 Q6UWX4 HHIP-like protein 2 HHIPL2 Q6UWY0 Arylsulfatase K ARSK Q6UWY2 Serine protease 57 PRSS57 Q6UWY5 Olfactomedin-like protein 1 OLFML1 Q6UX06 Olfactomedin-4 OLFM4 Q6UX07 Dehydrogenase/reductase SDR family member 13 DHRS13 Q6UX39 Amelotin AMTN Q6UX46 Protein FAM150B FAM150B Q6UX73 UPF0764 protein C16orf89 C16orf89 Q6UXB0 Protein FAM131A FAM131A Q6UXB1 Insulin growth factor-like family member 3 IGFL3 Q6UXB2 VEGF co-regulated chemokine 1 CXCL17 Q6UXF7 C-type lectin domain family 18 member B CLEC18B Q6UXH0 Hepatocellular carcinoma-associated protein TD26 C19orf80 Q6UXH1 Cysteine-rich with EGF-like domain protein 2 CRELD2 Q6UXH8 Collagen and calcium-binding EGF domain- CCBE1 containing protein 1 Q6UXH9 Inactive serine protease PAMR1 PAMR1 Q6UXI7 Vitrin VIT Q6UXI9 Nephronectin NPNT Q6UXN2 Trem-like transcript 4 protein TREML4 Q6UXS0 C-type lectin domain family 19 member A CLEC19A Q6UXT8 Protein FAM150A FAM150A Q6UXT9 Abhydrolase domain-containing protein 15 ABHD15 Q6UXV4 Apolipoprotein O-like APOOL Q6UXX5 Inter-alpha-trypsin inhibitor heavy chain H6 ITIH6 Q6UXX9 R-spondin-2 RSPO2 Q6UY14 ADAMTS-like protein 4 ADAMTSL4 Q6UY27 Prostate and testis expressed protein 2 PATE2 Q6W4X9 Mucin-6 MUC6 Q6WN34 Chordin-like protein 2 CHRDL2 Q6WRI0 Immunoglobulin superfamily member 10 IGSF10 Q6X4U4 Sclerostin domain-containing protein 1 SOSTDC1 Q6X784 Zona pellucida-binding protein 2 ZPBP2 Q6XE38 Secretoglobin family 1D member 4 SCGB1D4 Q6XPR3 Repetin RPTN Q6XZB0 Lipase member I LIPI Q6ZMM2 ADAMTS-like protein 5 ADAMTSL5 Q6ZMP0 Thrombospondin type-1 domain-containing THSD4 protein 4 Q6ZNF0 Iron/zinc purple acid phosphatase-like protein PAPL Q6ZRI0 Otogelin OTOG Q6ZRP7 Sulfhydryl oxidase 2 QSOX2 Q6ZWJ8 Kielin/chordin-like protein KCP Q75N90 Fibrillin-3 FBN3 Q765I0 Urotensin-2B UTS2D Q76B58 Protein FAM5C FAM5C Q76LX8 A disintegrin and metalloproteinase with ADAMTS13 thrombospondin motifs 13 Q76M96 Coiled-coil domain-containing protein 80 CCDC80 Q7L1S5 Carbohydrate sulfotransferase 9 CHST9 Q7L513 Fc receptor-like A FCRLA Q7L8A9 Vasohibin-1 VASH1 Q7RTM1 Otopetrin-1 OTOP1 Q7RTW8 Otoancorin OTOA Q7RTY5 Serine protease 48 PRSS48 Q7RTY7 Ovochymase-1 OVCH1 Q7RTZ1 Ovochymase-2 OVCH2 Q7Z304 MAM domain-containing protein 2 MAMDC2 Q7Z3S9 Notch homolog 2 N-terminal-like protein NOTCH2NL Q7Z4H4 Intermedin-short ADM2 Q7Z4P5 Growth/differentiation factor 7 GDF7 Q7Z4R8 UPF0669 protein C6orf120 C6orf120 Q7Z4W2 Lysozyme-like protein 2 LYZL2 Q7Z5A4 Serine protease 42 PRSS42 Q7Z5A7 Protein FAM19A5 FAM19A5 Q7Z5A8 Protein FAM19A3 FAM19A3 Q7Z5A9 Protein FAM19A1 FAM19A1 Q7Z5J1 Hydroxysteroid 1-beta-dehydrogenase 1-like HSD11B1L protein Q7Z5L0 Vitelline membrane outer layer protein 1 homolog VMO1 Q7Z5L3 Complement C1q-like protein 2 C1QL2 Q7Z5L7 Podocan PODN Q7Z5P4 17-beta-hydroxysteroid dehydrogenase 13 HSD17B13 Q7Z5P9 Mucin-19 MUC19 Q7Z5Y6 Bone morphogenetic protein 8A BMP8A Q7Z7B7 Beta-defensin 132 DEFB132 Q7Z7B8 Beta-defensin 128 DEFB128 Q7Z7C8 Transcription initiation factor TFIID subunit 8 TAF8 Q7Z7H5 Transmembrane emp24 domain-containing protein 4 TMED4 Q86SG7 Lysozyme g-like protein 2 LYG2 Q86SI9 Protein CEI C5orf38 Q86TE4 Leucine zipper protein 2 LUZP2 Q86TH1 ADAMTS-like protein 2 ADAMTSL2 Q86U17 Serpin A11 SERPINA11 Q86UU9 Endokinin-A TAC4 Q86UW8 Hyaluronan and proteoglycan link protein 4 HAPLN4 Q86UX2 Inter-alpha-trypsin inhibitor heavy chain H5 ITIH5 Q86V24 Adiponectin receptor protein 2 ADIPOR2 Q86VB7 Soluble CD 163 CD163 Q86VR8 Four-jointed box protein 1 FJX1 Q86WD7 Serpin A9 SERPINA9 Q86WN2 Interferon epsilon IFNE Q86WS3 Placenta-specific 1-like protein PLAC1L Q86X52 Chondroitin sulfate synthase 1 CHSY1 Q86XP6 Gastrokine-2 GKN2 Q86XS5 Angiopoietin-related protein 5 ANGPTL5 Q86Y27 B melanoma antigen 5 BAGE5 Q86Y28 B melanoma antigen 4 BAGE4 Q86Y29 B melanoma antigen 3 BAGE3 Q86Y30 B melanoma antigen 2 BAGE2 Q86Y38 Xylosyltransferase 1 XYLT1 Q86Y78 Ly6/PLAUR domain-containing protein 6 LYPD6 Q86YD3 Transmembrane protein 25 TMEM25 Q86YJ6 Threonine synthase-like 2 THNSL2 Q86YW7 Glycoprotein hormone beta-5 GPHB5 Q86Z23 Complement C1q-like protein 4 C1QL4 Q8IU57 Interleukin-28 receptor subunit alpha IL28RA Q8IUA0 WAP four-disulfide core domain protein 8 WFDC8 Q8IUB2 WAP four-disulfide core domain protein 3 WFDC3 Q8IUB3 Protein WFDC10B WFDC10B Q8IUB5 WAP four-disulfide core domain protein 13 WFDC13 Q8IUH2 Protein CREG2 CREG2 Q8IUK5 Plexin domain-containing protein 1 PLXDC1 Q8IUL8 Cartilage intermediate layer protein 2 C2 CILP2 Q8IUX7 Adipocyte enhancer-binding protein 1 AEBP1 Q8IUX8 Epidermal growth factor-like protein 6 EGFL6 Q8IVL8 Carboxypeptidase O CPO Q8IVN8 Somatomedin-B and thrombospondin type-1 SBSPON domain-containing protein Q8IVW8 Protein spinster homolog 2 SPNS2 Q8IW75 Serpin A12 SERPINA12 Q8IW92 Beta-galactosidase-1-like protein 2 GLB1L2 Q8IWL1 Pulmonary surfactant-associated protein A2 SFTPA2 Q8IWL2 Pulmonary surfactant-associated protein A1 SFTPA1 Q8IWV2 Contactin-4 CNTN4 Q8IWY4 Signal peptide, CUB and EGF-like domain- SCUBE1 containing protein 1 Q8IX30 Signal peptide, CUB and EGF-like domain- SCUBE3 containing protein 3 Q8IXA5 Sperm acrosome membrane-associated protein 3, SPACA3 membrane form Q8IXB1 DnaJ homolog subfamily C member 10 DNAJC10 Q8IXL6 Extracellular serine/threonine protein kinase FAM20C Fam20C Q8IYD9 Lung adenoma susceptibility protein 2 LAS2 Q8IYP2 Serine protease 58 PRSS58 Q8IYS5 Osteoclast-associated immunoglobulin-like OSCAR receptor Q8IZC6 Collagen alpha-1(XXVII) chain COL27A1 Q8IZJ3 C3 and PZP-like alpha-2-macroglobulin domain- CPAMD8 containing protein 8 Q8IZN7 Beta-defensin 107 DEFB107B Q8N0V4 Leucine-rich repeat LGI family member 2 LGI2 Q8N104 Beta-defensin 106 DEFB106B Q8N119 Matrix metalloproteinase-21 MMP21 Q8N129 Protein canopy homolog 4 CNPY4 Q8N135 Leucine-rich repeat LGI family member 4 LGI4 Q8N145 Leucine-rich repeat LGI family member 3 LGI3 Q8N158 Glypican-2 GPC2 Q8N1E2 Lysozyme g-like protein 1 LYG1 Q8N2E2 von Willebrand factor D and EGF domain- VWDE containing protein Q8N2E6 Prosalusin TOR2A Q8N2S1 Latent-transforming growth factor beta-binding LTBP4 protein 4 Q8N302 Angiogenic factor with G patch and FHA domains 1 AGGF1 Q8N307 Mucin-20 MUC20 Q8N323 NXPE family member 1 NXPE1 Q8N387 Mucin-15 MUC15 Q8N3Z0 Inactive serine protease 35 PRSS35 Q8N436 Inactive carboxypeptidase-like protein X2 CPXM2 Q8N474 Secreted frizzled-related protein 1 SFRP1 Q8N475 Follistatin-related protein 5 FSTL5 Q8N4F0 BPI fold-containing family B member 2 BPIFB2 Q8N4T0 Carboxypeptidase A6 CPA6 Q8N5W8 Protein FAM24B FAM24B Q8N687 Beta-defensin 125 DEFB125 Q8N688 Beta-defensin 123 DEFB123 Q8N690 Beta-defensin 119 DEFB119 Q8N6C5 Immunoglobulin superfamily member 1 IGSF1 Q8N6C8 Leukocyte immunoglobulin-like receptor LILRA3 subfamily A member 3 Q8N6G6 ADAMTS-like protein 1 ADAMTSL1 Q8N6Y2 Leucine-rich repeat-containing protein 17 LRRC17 Q8N729 Neuropeptide W-23 NPW Q8N8U9 BMP-binding endothelial regulator protein BMPER Q8N907 DAN domain family member 5 DAND5 Q8NAT1 Glycosyltransferase-like domain-containing GTDC2 protein 2 Q8NAU1 Fibronectin type III domain-containing protein 5 FNDC5 Q8NB37 Parkinson disease 7 domain-containing protein 1 PDDC1 Q8NBI3 Draxin DRAXIN Q8NBM8 Prenylcysteine oxidase-like PCYOX1L Q8NBP7 Proprotein convertase subtilisin/kexin type 9 PCSK9 Q8NBQ5 Estradiol 17-beta-dehydrogenase 11 HSD17B11 Q8NBV8 Synaptotagmin-8 SYT8 Q8NCC3 Group XV phospholipase A2 PLA2G15 Q8NCF0 C-type lectin domain family 18 member C CLEC18C Q8NCW5 NAD(P)H-hydrate epimerase APOA1BP Q8NDA2 Hemicentin-2 HMCN2 Q8NDX9 Lymphocyte antigen 6 complex locus protein G5b LY6G5B Q8NDZ4 Deleted in autism protein 1 C3orf58 Q8NEB7 Acrosin-binding protein ACRBP Q8NES8 Beta-defensin 124 DEFB124 Q8NET1 Beta-defensin 108B DEFB108B Q8NEX5 Protein WFDC9 WFDC9 Q8NEX6 Protein WFDC11 WFDC11 Q8NF86 Serine protease 33 PRSS33 Q8NFM7 Interleukin-17 receptor D IL17RD Q8NFQ5 BPI fold-containing family B member 6 BPIFB6 Q8NFQ6 BPI fold-containing family C protein BPIFC Q8NFU4 Follicular dendritic cell secreted peptide FDCSP Q8NFW1 Collagen alpha-1(XXII) chain COL22A1 Q8NG35 Beta-defensin 105 DEFB105B Q8NG41 Neuropeptide B-23 NPB Q8NHW6 Otospiralin OTOS Q8NI99 Angiopoietin-related protein 6 ANGPTL6 Q8TAA1 Probable ribonuclease 11 RNASE11 Q8TAG5 V-set and transmembrane domain-containing VSTM2A protein 2A Q8TAL6 Fin bud initiation factor homolog FIBIN Q8TAT2 Fibroblast growth factor-binding protein 3 FGFBP3 Q8TAX7 Mucin-7 MUC7 Q8TB22 Spermatogenesis-associated protein 20 SPATA20 Q8TB73 Protein NDNF NDNF Q8TB96 T-cell immunomodulatory protein ITFG1 Q8TC92 Protein disulfide-thiol oxidoreductase ENOX1 Q8TCV5 WAP four-disulfide core domain protein 5 WFDC5 Q8TD06 Anterior gradient protein 3 homolog AGR3 Q8TD33 Secretoglobin family 1C member 1 SCGB1C1 Q8TD46 Cell surface glycoprotein CD200 receptor 1 CD200R1 Q8TDE3 Ribonuclease 8 RNASE8 Q8TDF5 Neuropilin and tolloid-like protein 1 NETO1 Q8TDL5 BPI fold-containing family B member 1 BPIFB1 Q8TE56 A disintegrin and metalloproteinase with ADAMTS17 thrombospondin motifs 17 Q8TE57 A disintegrin and metalloproteinase with ADAMTS16 thrombospondin motifs 16 Q8TE58 A disintegrin and metalloproteinase with ADAMTS15 thrombospondin motifs 15 Q8TE59 A disintegrin and metalloproteinase with ADAMTS19 thrombospondin motifs 19 Q8TE60 A disintegrin and metalloproteinase with ADAMTS18 thrombospondin motifs 18 Q8TE99 Acid phosphatase-like protein 2 ACPL2 Q8TER0 Sushi, nidogen and EGF-like domain-containing SNED1 protein 1 Q8TEU8 WAP, kazal, immunoglobulin, kunitz and NTR WFIKKN2 domain-containing protein 2 Q8WTQ1 Beta-defensin 104 DEFB104B Q8WTR8 Netrin-5 NTN5 Q8WTU2 Scavenger receptor cysteine-rich domain- SRCRB4D containing group B protein Q8WU66 Protein TSPEAR TSPEAR Q8WUA8 Tsukushin TSKU Q8WUF8 Protein FAM172A FAM172A Q8WUJ1 Neuferricin CYB5D2 Q8WUY1 UPF0670 protein THEM6 THEM6 Q8WVN6 Secreted and transmembrane protein 1 SECTM1 Q8WVQ1 Soluble calcium-activated nucleotidase 1 CANT1 Q8WWA0 Intelectin-1 ITLN1 Q8WWG1 Neuregulin-4 NRG4 Q8WWQ2 Inactive heparanase-2 HPSE2 Q8WWU7 Intelectin-2 ITLN2 Q8WWY7 WAP four-disulfide core domain protein 12 WFDC12 Q8WWY8 Lipase member H LIPH Q8WWZ8 Oncoprotein-induced transcript 3 protein OIT3 Q8WX39 Epididymal-specific lipocalin-9 LCN9 Q8WXA2 Prostate and testis expressed protein 1 PATE1 Q8WXD2 Secretogranin-3 SCG3 Q8WXF3 Relaxin-3 A chain RLN3 Q8WXI7 Mucin-16 MUC16 Q8WXQ8 Carboxypeptidase A5 CPA5 Q8WXS8 A disintegrin and metalloproteinase with ADAMTS14 thrombospondin motifs 14 Q92484 Acid sphingomyelinase-like phosphodiesterase 3a SMPDL3A Q92485 Acid sphingomyelinase-like phosphodiesterase 3b SMPDL3B Q92496 Complement factor H-related protein 4 CFHR4 Q92520 Protein FAM3C FAM3C Q92563 Testican-2 SPOCK2 Q92583 C-C motif chemokine 17 CCL17 Q92626 Peroxidasin homolog PXDN Q92743 Serine protease HTRA1 HTRA1 Q92752 Tenascin-R TNR Q92765 Secreted frizzled-related protein 3 FRZB Q92819 Hyaluronan synthase 2 HAS2 Q92820 Gamma-glutamyl hydrolase GGH Q92824 Proprotein convertase subtilisin/kexin type 5 PCSK5 Q92832 Protein kinase C-binding protein NELL1 NELL1 Q92838 Ectodysplasin-A, membrane form EDA Q92874 Deoxyribonuclease-1-like 2 DNASE1L2 Q92876 Kallikrein-6 KLK6 Q92913 Fibroblast growth factor 13 FGF13 Q92954 Proteoglycan 4 C-terminal part PRG4 Q93038 Tumor necrosis factor receptor superfamily TNFRSF25 member 25 Q93091 Ribonuclease K6 RNASE6 Q93097 Protein Wnt-2b WNT2B Q93098 Protein Wnt-8b WNT8B Q95460 Major histocompatibility complex class I-related MR1 gene protein Q969D9 Thymic stromal lymphopoietin TSLP Q969E1 Liver-expressed antimicrobial peptide 2 LEAP2 Q969H8 UPF0556 protein C19orf10 C19orf10 Q969Y0 NXPE family member 3 NXPE3 Q96A54 Adiponectin receptor protein 1 ADIPOR1 Q96A83 Collagen alpha-1(XXVI) chain EMID2 Q96A84 EMI domain-containing protein 1 EMID1 Q96A98 Tuberoinfundibular peptide of 39 residues PTH2 Q96A99 Pentraxin-4 PTX4 Q96BH3 Epididymal sperm-binding protein 1 ELSPBP1 Q96BQ1 Protein FAM3D FAM3D Q96CG8 Collagen triple helix repeat-containing protein 1 CTHRC1 Q96DA0 Zymogen granule protein 16 homolog B ZG16B Q96DN2 von Willebrand factor C and EGF domain- VWCE containing protein Q96DR5 BPI fold-containing family A member 2 BPIFA2 Q96DR8 Mucin-like protein 1 MUCL1 Q96DX4 RING finger and SPRY domain-containing protein 1 RSPRY1 Q96EE4 Coiled-coil domain-containing protein 126 CCDC126 Q96GS6 Abhydrolase domain-containing protein FAM108A1 FAM108A1 Q96GW7 Brevican core protein BCAN Q96HF1 Secreted frizzled-related protein 2 SFRP2 Q96I82 Kazal-type serine protease inhibitor domain- KAZALD1 containing protein 1 Q96ID5 Immunoglobulin superfamily member 21 IGSF21 Q96II8 Leucine-rich repeat and calponin homology LRCH3 domain-containing protein 3 Q96IY4 Carboxypeptidase B2 CPB2 Q96JB6 Lysyl oxidase homolog 4 LOXL4 Q96JK4 HHIP-like protein 1 HHIPL1 Q96KN2 Beta-Ala-His dipeptidase CNDP1 Q96KW9 Protein SPACA7 SPACA7 Q96KX0 Lysozyme-like protein 4 LYZL4 Q96L15 Ecto-ADP-ribosyltransferase 5 ART5 Q96LB8 Peptidoglycan recognition protein 4 PGLYRP4 Q96LB9 Peptidoglycan recognition protein 3 PGLYRP3 Q96LC7 Sialic acid-binding Ig-like lectin 10 SIGLEC10 Q96LR4 Protein FAM19A4 FAM19A4 Q96MK3 Protein FAM20A FAM20A Q96MS3 Glycosyltransferase 1 domain-containing protein 1 GLT1D1 Q96NY8 Processed poliovirus receptor-related protein 4 PVRL4 Q96NZ8 WAP, kazal, immunoglobulin, kunitz and NTR WFIKKN1 domain-containing protein 1 Q96NZ9 Proline-rich acidic protein 1 PRAP1 Q96P44 Collagen alpha-1(XXI) chain COL21A1 Q96PB7 Noelin-3 OLFM3 Q96PC5 Melanoma inhibitory activity protein 2 MIA2 Q96PD5 N-acetylmuramoyl-L-alanine amidase PGLYRP2 Q96PH6 Beta-defensin 118 DEFB118 Q96PL1 Secretoglobin family 3A member 2 SCGB3A2 Q96PL2 Beta-tectorin TECTB Q96QH8 Sperm acrosome-associated protein 5 SPACA5 Q96QR1 Secretoglobin family 3A member 1 SCGB3A1 Q96QU1 Protocadherin-15 PCDH15 Q96QV1 Hedgehog-interacting protein HHIP Q96RW7 Hemicentin-1 HMCN1 Q96S42 Nodal homolog NODAL Q96S86 Hyaluronan and proteoglycan link protein 3 HAPLN3 Q96SL4 Glutathione peroxidase 7 GPX7 Q96SM3 Probable carboxypeptidase X1 CPXM1 Q96T91 Glycoprotein hormone alpha-2 GPHA2 Q99062 Granulocyte colony-stimulating factor receptor CSF3R Q99102 Mucin-4 alpha chain MUC4 Q99217 Amelogenin, X isoform AMELX Q99218 Amelogenin, Y isoform AMELY Q99435 Protein kinase C-binding protein NELL2 NELL2 Q99470 Stromal cell-derived factor 2 SDF2 Q99542 Matrix metalloproteinase-19 MMP19 Q99574 Neuroserpin SERPINI1 Q99584 Protein S100-A13 S100A13 Q99616 C-C motif chemokine 13 CCL13 Q99645 Epiphycan EPYC Q99674 Cell growth regulator with EF hand domain CGREF1 protein 1 Q99715 Collagen alpha-1(XII) chain COL12A1 Q99727 Metalloproteinase inhibitor 4 TIMP4 Q99731 C-C motif chemokine 19 CCL19 Q99748 Neurturin NRTN Q99935 Proline-rich protein 1 PROL1 Q99942 E3 ubiquitin-protein ligase RNF5 RNF5 Q99944 Epidermal growth factor-like protein 8 EGFL8 Q99954 Submaxillary gland androgen-regulated protein 3A SMR3A Q99969 Retinoic acid receptor responder protein 2 RARRES2 Q99972 Myocilin MYOC Q99983 Osteomodulin OMD Q99985 Semaphorin-3C SEMA3C Q99988 Growth/differentiation factor 15 GDF15 Q9BPW4 Apolipoprotein L4 APOL4 Q9BQ08 Resistin-like beta RETNLB Q9BQ16 Testican-3 SPOCK3 Q9BQ51 Programmed cell death 1 ligand 2 PDCD1LG2 Q9BQB4 Sclerostin SOST Q9BQI4 Coiled-coil domain-containing protein 3 CCDC3 Q9BQP9 BPI fold-containing family A member 3 BPIFA3 Q9BQR3 Serine protease 27 PRSS27 Q9BQY6 WAP four-disulfide core domain protein 6 WFDC6 Q9BRR6 ADP-dependent glucokinase ADPGK Q9BS86 Zona pellucida-binding protein 1 ZPBP Q9BSG0 Protease-associated domain-containing protein 1 PRADC1 Q9BSG5 Retbindin RTBDN Q9BT30 Probable alpha-ketoglutarate-dependent ALKBH7 dioxygenase ABH7 Q9BT56 Spexin C12orG9 Q9BT67 NEDD4 family-interacting protein 1 NDFIP1 Q9BTY2 Plasma alpha-L-fucosidase FUCA2 Q9BU40 Chordin-like protein 1 CHRDL1 Q9BUD6 Spondin-2 SPON2 Q9BUN1 Protein MENT MENT Q9BUR5 Apolipoprotein O APOO Q9BV94 ER degradation-enhancing alpha-mannosidase-like 2 EDEM2 Q9BWP8 Collectin-11 COLEC11 Q9BWS9 Chitinase domain-containing protein 1 CHID1 Q9BX67 Junctional adhesion molecule C JAM3 Q9BX93 Group XIIB secretory phospholipase A2-like PLA2G12B protein Q9BXI9 Complement C1q tumor necrosis factor-related C1QTNF6 protein 6 Q9BXJ0 Complement C1q tumor necrosis factor-related C1QTNF5 protein 5 Q9BXJ1 Complement C1q tumor necrosis factor-related C1QTNF1 protein 1 Q9BXJ2 Complement C1q tumor necrosis factor-related C1QTNF7 protein 7 Q9BXJ3 Complement C1q tumor necrosis factor-related C1QTNF4 protein 4 Q9BXJ4 Complement C1q tumor necrosis factor-related C1QTNF3 protein 3 Q9BXJ5 Complement C1q tumor necrosis factor-related C1QTNF2 protein 2 Q9BXN1 Asporin ASPN Q9BXP8 Pappalysin-2 PAPPA2 Q9BXR6 Complement factor H-related protein 5 CFHR5 Q9BXS0 Collagen alpha-1(XXV) chain COL25A1 Q9BXX0 EMILIN-2 EMILIN2 Q9BXY4 R-spondin-3 RSPO3 Q9BY15 EGF-like module-containing mucin-like hormone EMR3 receptor-like 3 subunit beta Q9BY50 Signal peptidase complex catalytic subunit SEC11C SEC11C Q9BY76 Angiopoietin-related protein 4 ANGPTL4 Q9BYF1 Processed angiotensin-converting enzyme 2 ACE2 Q9BYJ0 Fibroblast growth factor-binding protein 2 FGFBP2 Q9BYW3 Beta-defensin 126 DEFB126 Q9BYX4 Interferon-induced helicase C domain-containing IFIH1 protein 1 Q9BYZ8 Regenerating islet-derived protein 4 REG4 Q9BZ76 Contactin-associated protein-like 3 CNTNAP3 Q9BZG9 Ly-6/neurotoxin-like protein 1 LYNX1 Q9BZJ3 Tryptase delta TPSD1 Q9BZM1 Group XIIA secretory phospholipase A2 PLA2G12A Q9BZM2 Group IIF secretory phospholipase A2 PLA2G2F Q9BZM5 NKG2D ligand 2 ULBP2 Q9BZP6 Acidic mammalian chitinase CHIA Q9BZZ2 Sialoadhesin SIGLEC1 Q9C0B6 Protein FAM5B FAM5B Q9GZM7 Tubulointerstitial nephritis antigen-like TINAGL1 Q9GZN4 Brain-specific serine protease 4 PRSS22 Q9GZP0 Platelet-derived growth factor D, receptor-binding PDGFD form Q9GZT5 Protein Wnt-10a WNT10A Q9GZU5 Nyctalopin NYX Q9GZV7 Hyaluronan and proteoglycan link protein 2 HAPLN2 Q9GZV9 Fibroblast growth factor 23 FGF23 Q9GZX9 Twisted gastrulation protein homolog 1 TWSG1 Q9GZZ7 GDNF family receptor alpha-4 GFRA4 Q9GZZ8 Extracellular glycoprotein lacritin LACRT Q9H0B8 Cysteine-rich secretory protein LCCL domain- CRISPLD2 containing 2 Q9H106 Signal-regulatory protein delta SIRPD Q9H114 Cystatin-like 1 CSTL1 Q9H173 Nucleotide exchange factor SIL1 SIL1 Q9H1E1 Ribonuclease 7 RNASE7 Q9H1F0 WAP four-disulfide core domain protein 10A WFDC10A Q9H1J5 Protein Wnt-8a WNT8A Q9H1J7 Protein Wnt-5b WNT5B Q9H1M3 Beta-defensin 129 DEFB129 Q9H1M4 Beta-defensin 127 DEFB127 Q9H1Z8 Augurin C2orf40 Q9H239 Matrix metalloproteinase-28 MMP28 Q9H2A7 C-X-C motif chemokine 16 CXCL16 Q9H2A9 Carbohydrate sulfotransferase 8 CHST8 Q9H2R5 Kallikrein-15 KLK15 Q9H2X0 Chordin CHRD Q9H2X3 C-type lectin domain family 4 member M CLEC4M Q9H306 Matrix metalloproteinase-27 MMP27 Q9H324 A disintegrin and metalloproteinase with ADAMTS10 thrombospondin motifs 10 Q9H336 Cysteine-rich secretory protein LCCL domain- CRISPLD1 containing 1 Q9H3E2 Sorting nexin-25 SNX25 Q9H3R2 Mucin-13 MUC13 Q9H3U7 SPARC-related modular calcium-binding protein 2 SMOC2 Q9H3Y0 Peptidase inhibitor R3HDML R3HDML Q9H4A4 Aminopeptidase B RNPEP Q9H4F8 SPARC-related modular calcium-binding protein 1 SMOC1 Q9H4G1 Cystatin-9-like CST9L Q9H5V8 CUB domain-containing protein 1 CDCP1 Q9H6B9 Epoxide hydrolase 3 EPHX3 Q9H6E4 Coiled-coil domain-containing protein 134 CCDC134 Q9H741 UPF0454 protein C12orf49 C12orf49 Q9H772 Gremlin-2 GREM2 Q9H7Y0 Deleted in autism-related protein 1 CXorf36 Q9H8L6 Multimerin-2 MMRN2 Q9H9S5 Fukutin-related protein FKRP Q9HAT2 Sialate O-acetylesterase SIAE Q9HB40 Retinoid-inducible serine carboxypeptidase SCPEP1 Q9HB63 Netrin-4 NTN4 Q9HBJ0 Placenta-specific protein 1 PLAC1 Q9HC23 Prokineticin-2 PROK2 Q9HC57 WAP four-disulfide core domain protein 1 WFDC1 Q9HC73 Cytokine receptor-like factor 2 CRLF2 Q9HC84 Mucin-5B MUC5B Q9HCB6 Spondin-1 SPON1 Q9HCQ7 Neuropeptide NPSF NPVF Q9HCT0 Fibroblast growth factor 22 FGF22 Q9HD89 Resistin RETN Q9NNX1 Tuftelin TUFT1 Q9NNX6 CD209 antigen CD209 Q9NP55 BPI fold-containing family A member 1 BPIFA1 Q9NP70 Ameloblastin AMBN Q9NP95 Fibroblast growth factor 20 FGF20 Q9NP99 Triggering receptor expressed on myeloid cells 1 TREM1 Q9NPA2 Matrix metalloproteinase-25 MMP25 Q9NPE2 Neugrin NGRN Q9NPH0 Lysophosphatidic acid phosphatase type 6 ACP6 Q9NPH6 Odorant-binding protein 2b OBP2B Q9NQ30 Endothelial cell-specific molecule 1 ESM1 Q9NQ36 Signal peptide, CUB and EGF-like domain- SCUBE2 containing protein 2 Q9NQ38 Serine protease inhibitor Kazal-type 5 SPINK5 Q9NQ76 Matrix extracellular phosphoglycoprotein MEPE Q9NQ79 Cartilage acidic protein 1 CRTAC1 Q9NR16 Scavenger receptor cysteine-rich type 1 protein CD163L1 M160 Q9NR23 Growth/differentiation factor 3 GDF3 Q9NR71 Neutral ceramidase ASAH2 Q9NR99 Matrix-remodeling-associated protein 5 MXRA5 Q9NRA1 Platelet-derived growth factor C PDGFC Q9NRC9 Otoraplin OTOR Q9NRE1 Matrix metalloproteinase-26 MMP26 Q9NRJ3 C-C motif chemokine 28 CCL28 Q9NRM1 Enamelin ENAM Q9NRN5 Olfactomedin-like protein 3 OLFML3 Q9NRR1 Cytokine-like protein 1 CYTL1 Q9NS15 Latent-transforming growth factor beta-binding LTBP3 protein 3 Q9NS62 Thrombospondin type-1 domain-containing THSD1 protein 1 Q9NS71 Gastrokine-1 GKN1 Q9NS98 Semaphorin-3G SEMA3G Q9NSA1 Fibroblast growth factor 21 FGF21 Q9NT22 EMILIN-3 EMILIN3 Q9NTU7 Cerebellin-4 CBLN4 Q9NVR0 Kelch-like protein 11 KLHL11 Q9NWH7 Spermatogenesis-associated protein 6 SPATA6 Q9NXC2 Glucose-fructose oxidoreductase domain- GFOD1 containing protein 1 Q9NY56 Odorant-binding protein 2a OBP2A Q9NY84 Vascular non-inflammatory molecule 3 VNN3 Q9NZ20 Group 3 secretory phospholipase A2 PLA2G3 Q9NZC2 Triggering receptor expressed on myeloid cells 2 TREM2 Q9NZK5 Adenosine deaminase CECR1 CECR1 Q9NZK7 Group IIE secretory phospholipase A2 PLA2G2E Q9NZP8 Complement C1r subcomponent-like protein C1RL Q9NZV1 Cysteine-rich motor neuron 1 protein CRIM1 Q9NZW4 Dentin sialoprotein DSPP Q9P0G3 Kallikrein-14 KLK14 Q9P0W0 Interferon kappa IFNK Q9P218 Collagen alpha-1(XX) chain COL20A1 Q9P2C4 Transmembrane protein 181 TMEM181 Q9P2K2 Thioredoxin domain-containing protein 16 TXNDC16 Q9P2N4 A disintegrin and metalloproteinase with ADAMTS9 thrombospondin motifs 9 Q9UBC7 Galanin-like peptide GALP Q9UBD3 Cytokine SCM-1 beta XCL2 Q9UBD9 Cardiotrophin-like cytokine factor 1 CLCF1 Q9UBM4 Opticin OPTC Q9UBP4 Dickkopf-related protein 3 DKK3 Q9UBQ6 Exostosin-like 2 EXTL2 Q9UBR5 Chemokine-like factor CKLF Q9UBS5 Gamma-aminobutyric acid type B receptor subunit GABBR1 1 Q9UBT3 Dickkopf-related protein 4 short form DKK4 Q9UBU2 Dickkopf-related protein 2 DKK2 Q9UBU3 Ghrelin-28 GHRL Q9UBV4 Protein Wnt-16 WNT16 Q9UBX5 Fibulin-5 FBLN5 Q9UBX7 Kallikrein-11 KLK11 Q9UEF7 Klotho KL Q9UFP1 Protein FAM198A FAM198A Q9UGM3 Deleted in malignant brain tumors 1 protein DMBT1 Q9UGM5 Fetuin-B FETUB Q9UGP8 Translocation protein SEC63 homolog SEC63 Q9UHF0 Neurokinin-B TAC3 Q9UHF1 Epidermal growth factor-like protein 7 EGFL7 Q9UHG2 ProSAAS PCSK1N Q9UHI8 A disintegrin and metalloproteinase with ADAMTS1 thrombospondin motifs 1 Q9UHL4 Dipeptidyl peptidase 2 DPP7 Q9UI42 Carboxypeptidase A4 CPA4 Q9UIG4 Psoriasis susceptibility 1 candidate gene 2 protein PSORS1C2 Q9UIK5 Tomoregulin-2 TMEFF2 Q9UIQ6 Leucyl-cystinyl aminopeptidase, pregnancy serum LNPEP form Q9UJA9 Ectonucleotide ENPP5 pyrophosphatase/phosphodiesterase family member 5 Q9UJH8 Meteorin METRN Q9UJJ9 N-acetylglucosamine-1-phosphotransferase GNPTG subunit gamma Q9UJW2 Tubulointerstitial nephritis antigen TINAG Q9UK05 Growth/differentiation factor 2 GDF2 Q9UK55 Protein Z-dependent protease inhibitor SERPINA10 Q9UK85 Dickkopf-like protein 1 DKKL1 Q9UKJ1 Paired immunoglobulin-like type 2 receptor alpha PILRA Q9UKP4 A disintegrin and metalloproteinase with ADAMTS7 thrombospondin motifs 7 Q9UKP5 A disintegrin and metalloproteinase with ADAMTS6 thrombospondin motifs 6 Q9UKQ2 Disintegrin and metalloproteinase domain- ADAM28 containing protein 28 Q9UKQ9 Kallikrein-9 KLK9 Q9UKR0 Kallikrein-12 KLK12 Q9UKR3 Kallikrein-13 KLK13 Q9UKU9 Angiopoietin-related protein 2 ANGPTL2 Q9UKZ9 Procollagen C-endopeptidase enhancer 2 PCOLCE2 Q9UL52 Transmembrane protease serine 11E non-catalytic TMPRSS11E chain Q9ULC0 Endomucin EMCN Q9ULI3 Protein HEG homolog 1 HEG1 Q9ULZ1 Apelin-13 APLN Q9ULZ9 Matrix metalloproteinase-17 MMP17 Q9UM21 Alpha-1,3-mannosyl-glycoprotein 4-beta-N- MGAT4A acetylglucosaminyltransferase A soluble form Q9UM22 Mammalian ependymin-related protein 1 EPDR1 Q9UM73 ALK tyrosine kinase receptor ALK Q9UMD9 97 kDa linear IgA disease antigen COL17A1 Q9UMX5 Neudesin NENF Q9UN73 Protocadherin alpha-6 PCDHA6 Q9UNA0 A disintegrin and metalloproteinase with ADAMTS5 thrombospondin motifs 5 Q9UNI1 Chymotrypsin-like elastase family member 1 CELA1 Q9UNK4 Group IID secretory phospholipase A2 PLA2G2D Q9UP79 A disintegrin and metalloproteinase with ADAMTS8 thrombospondin motifs 8 Q9UPZ6 Thrombospondin type-1 domain-containing THSD7A protein 7A Q9UQ72 Pregnancy-specific beta-1-glycoprotein 11 PSG11 Q9UQ74 Pregnancy-specific beta-1-glycoprotein 8 PSG8 Q9UQC9 Calcium-activated chloride channel regulator 2 CLCA2 Q9UQE7 Structural maintenance of chromosomes protein 3 SMC3 Q9UQP3 Tenascin-N TNN Q9Y223 UDP-N-acetylglucosamine 2-epimerase GNE Q9Y240 C-type lectin domain family 11 member A CLEC11A Q9Y251 Heparanase 8 kDa subunit HPSE Q9Y258 C-C motif chemokine 26 CCL26 Q9Y264 Angiopoietin-4 ANGPT4 Q9Y275 Tumor necrosis factor ligand superfamily member TNFSF13B 13b, membrane form Q9Y287 BRI2 intracellular domain ITM2B Q9Y2E5 Epididymis-specific alpha-mannosidase MAN2B2 Q9Y334 von Willebrand factor A domain-containing VWA7 protein 7 Q9Y337 Kallikrein-5 KLK5 Q9Y3B3 Transmembrane emp24 domain-containing protein 7 TMED7 Q9Y3E2 BolA-like protein 1 BOLA1 Q9Y426 C2 domain-containing protein 2 C2CD2 Q9Y4K0 Lysyl oxidase homolog 2 LOXL2 Q9Y4X3 C-C motif chemokine 27 CCL27 Q9Y5C1 Angiopoietin-related protein 3 ANGPTL3 Q9Y5I2 Protocadherin alpha-10 PCDHA10 Q9Y5I3 Protocadherin alpha-1 PCDHA1 Q9Y5K2 Kallikrein-4 KLK4 Q9Y5L2 Hypoxia-inducible lipid droplet-associated protein HILPDA Q9Y5Q5 Atrial natriuretic peptide-converting enzyme CORIN Q9Y5R2 Matrix metalloproteinase-24 MMP24 Q9Y5U5 Tumor necrosis factor receptor superfamily TNFRSF18 member 18 Q9Y5W5 Wnt inhibitory factor 1 WIF1 Q9Y5X9 Endothelial lipase LIPG Q9Y625 Secreted glypican-6 GPC6 Q9Y646 Carboxypeptidase Q CPQ Q9Y6C2 EMILIN-1 EMILIN1 Q9Y6F9 Protein Wnt-6 WNT6 Q9Y6I9 Testis-expressed sequence 264 protein TEX264 Q9Y6L7 Tolloid-like protein 2 TLL2 Q9Y6N3 Calcium-activated chloride channel regulator CLCA3P family member 3 Q9Y6N6 Laminin subunit gamma-3 LAMC3 Q9Y6R7 IgGFc-binding protein FCGBP Q9Y6Y9 Lymphocyte antigen 96 LY96 Q9Y6Z7 Collectin-10 COLEC10

In some embodiments, the compositions and methods of the invention provide for the delivery of one or more mRNAs encoding one or more additional exemplary proteins listed in Table 2; thus, compositions of the invention may comprise an mRNA encoding a protein listed in Table 2 (or a homolog thereof) along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein chosen from the proteins listed in Table 2 (or a homolog thereof) along with other components set out herein.

In some embodiments, compositions and methods of the invention provide for the delivery of one or more nucleic acids that bind to or otherwise act on an endogenous nucleic acid, such as endogenous mRNA or DNA, that alter the transcription, translation, secretion, or action of one or more proteins or genes listed in Table 2 (or a homolog thereof). For example, RNAi nucleic acids bind to endogenous mRNA, miRNA, or other RNAs, while guide RNA (gRNA) and CRISPR RNA (crRNA) respectively bind to DNA and act on DNA in the nucleus. Accordingly, in embodiments, the compositions and methods of the invention provide for delivery of nucleic acids including one or more of RNAi nucleic acids, gRNA and crRNA to interfere with the transcription, translation, secretion, or action of one or more proteins or genes listed in Table 2 (and homologs thereof) along with other components set out herein.

TABLE 2 Additional Exemplary Proteins Uniprot ID Protein Name Gene Name A6NGW2 Putative stereocilin-like protein STRCP1 A6NIE9 Putative serine protease 29 PRSS29P A6NJ16 Putative V-set and immunoglobulin IGHV4OR15-8 domain-containing-like protein IGHV4OR15-8 A6NJS3 Putative V-set and immunoglobulin IGHV1OR21-1 domain-containing-like protein IGHV1OR21-1 A6NMY6 Putative annexin A2-like protein ANXA2P2 A8MT79 Putative zinc-alpha-2-glycoprotein-like 1 A8MWS1 Putative killer cell immunoglobulin-like KIR3DP1 receptor like protein KIR3DP1 A8MXU0 Putative beta-defensin 108A DEFB108P1 C9JUS6 Putative adrenomedullin-5-like protein ADM5 P0C7V7 Putative signal peptidase complex SEC11B catalytic subunit SEC11B P0C854 Putative cat eye syndrome critical region CECR9 protein 9 Q13046 Putative pregnancy-specific beta-1- PSG7 glycoprotein 7 Q16609 Putative apolipoprotein(a)-like protein 2 LPAL2 Q2TV78 Putative macrophage-stimulating protein MST1P9 MSTP9 Q5JQD4 Putative peptide YY-3 PYY3 Q5R387 Putative inactive group IIC secretory PLA2G2C phospholipase A2 Q5VSP4 Putative lipocalin 1-like protein 1 LCN1P1 Q5W188 Putative cystatin-9-like protein CST9LP1 CST9LP1 Q6UXR4 Putative serpin A13 SERPINA13P Q86SH4 Putative testis-specific prion protein PRNT Q86YQ2 Putative latherin LATH Q8IVG9 Putative humanin peptide MT-RNR2 Q8NHM4 Putative trypsin-6 TRY6 Q8NHW4 C-C motif chemokine 4-like CCL4L2 Q9H7L2 Putative killer cell immunoglobulin-like KIR3DX1 receptor-like protein KIR3DX1 Q9NRI6 Putative peptide YY-2 PYY2 Q9UF72 Putative TP73 antisense gene protein 1 TP73-AS1 Q9UKY3 Putative inactive carboxylesterase 4 CES1P1

The Uniprot IDs set forth in Table 1 and Table 2 refer to the human versions the listed proteins and the sequences of each are available from the Uniprot database. Sequences of the listed proteins are also generally available for various animals, including various mammals and animals of veterinary or industrial interest. Accordingly, in some embodiments, compositions and methods of the invention provide for the delivery of one or more mRNAs encoding one or more proteins chosen from mammalian homologs or homologs from an animal of veterinary or industrial interest of the secreted proteins listed in Table 1 and Table 2; thus, compositions of the invention may comprise an mRNA encoding a protein chosen from mammalian homologs or homologs from an animal of veterinary or industrial interest of a protein listed in Table 1 and Table 2 along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein chosen from mammalian homologs or homologs from an animal of veterinary or industrial interest of a protein listed in Table 1 and Table 2 along with other components set out herein. In some embodiments, mammalian homologs are chosen from mouse, rat, hamster, gerbil, horse, pig, cow, llama, alpaca, mink, dog, cat, ferret, sheep, goat, or camel homologs. In some embodiments, the animal of veterinary or industrial interest is chosen from the mammals listed above and/or chicken, duck, turkey, salmon, catfish, or tilapia.

In embodiments, the compositions and methods of the invention provide for the delivery of mRNA encoding a protein chosen from Table 3. In some embodiments, the compositions and methods of the invention provide for the delivery of one or more mRNAs encoding one or more proteins listed in Table 3; thus, compositions of the invention may comprise an mRNA encoding a protein listed in Table 3 (or a homolog thereof) along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein chosen from the proteins listed in Table 3 (or a homolog thereof) along with other components set out herein.

In some embodiments, compositions and methods of the invention provide for the delivery of one or more nucleic acids that bind to or otherwise act on an endogenous nucleic acid, such as endogenous mRNA or DNA, that alter the transcription, translation, secretion, or action of one or more proteins listed in Table 3 (or a homolog thereof). For example, RNAi nucleic acids bind to endogenous mRNA, miRNA, or other RNAs, while guide RNA (gRNA) and CRISPR RNA (crRNA) respectively bind to DNA and act on DNA in the nucleus. Accordingly, in embodiments, the compositions and methods of the invention provide for delivery of nucleic acids including one or more of RNAi nucleic acids, gRNA and crRNA to interfere with the transcription, translation, secretion, or action of one or more proteins listed in Table 3 (and homologs thereof) along with other components set out herein.

TABLE 3 Lysosomal and Related Proteins α-fucosidase α-galactosidase α-glucosidase α-Iduronidase α-mannosidase α-N-acetylgalactosaminidase (α-galactosidase B) β-galactosidase β-glucuronidase β-hexosaminidase β-mannosidase 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) lyase 3-methylcrotonyl-CoA carboxylase 3-O-sulfogalactosyl cerebroside sulfatase (arylsulfatase A) acetyl-CoA transferase acid alpha-glucosidase acid ceramidase acid lipase acid phosphatase acid sphingomyelinase alpha-galactosidase A arylsulfatase A beta-galactosidase beta-glucocerebrosidase beta-hexosaminidase biotinidase cathepsin A cathepsin K CLN3 CLN5 CLN6 CLN8 CLN9 cystine transporter (cystinosin) cytosolic protein beta3A subunit of the adaptor protein-3 complex, AP3 formyl-Glycine generating enzyme (FGE) galactocerebrosidase galactose-1-phosphate uridyltransferase (GALT) galactose 6-sulfate sulfatase (also known as N-acetylgalactosamine-6-sulfatase) glucocerebrosidase glucuronate sulfatase glucuronidase glycoprotein cleaving enzymes glycosaminoglycan cleaving enzymes glycosylasparaginase (aspartylglucosaminidase) GM2-AP Heparan-alpha-glucosaminide N-acetyltransferase (HGSNAT, TMEM76) Heparan sulfatase hexosaminidase A lysosomal proteases methylmalonyl-CoA mutase hyaluronidase Iduronate sulfatase LAMP-2 lysosomal α-mannosidase Lysosomal p40 (C2orf18) Major facilitator superfamily domain containing 8 protein (MFSD8 or CLN7) N-acetylgalactosamine 4-sulfatase N-acetyl glucosamine 6-sulfatase N-acetyl glucosaminidase N-acetylglucosamine-1-phosphate transferase NPC1 NPC2 palmitoyl-protein thioesterase palmitoyl-protein thioesterase (CLN1) Saposin A (Sphingolipid activator protein A) Saposin B (Sphingolipid activator protein B) Saposin C (Sphingolipid activator protein C) Saposin D (Sphingolipid activator protein D) sialic acid transporter (sialin) sialidase Sialin sulfatase Transmembrane protein 74 (TMEM74) tripeptidyl-peptidase tripeptidyl-peptidase I (CLN2) UDP-N-acetylglucosamine-phosphotransferase

Information regarding lysosomal proteins is available from Lubke et al., “Proteomics of the Lysosome,” Biochim Biophys Acta. (2009) 1793: 625-635. In some embodiments, the protein listed in Table 3 and encoded by mRNA in the compositions and methods of the invention is a human protein. Sequences of the listed proteins are also available for various animals, including various mammals and animals of veterinary or industrial interest as described above.

In some embodiments, the compositions and methods of the invention provide for the delivery of mRNA encoding a therapeutic protein (e.g., cytosolic, transmembrane or secreted) such as those listed in Table 4. In some embodiments, the compositions and methods of the invention provide for the delivery of an mRNA encoding a therapeutic protein useful in treating a disease or disorder (i.e., indication) listed in Table 4; thus, compositions of the invention may comprise an mRNA encoding a therapeutic protein listed or not listed in Table 4 (or a homolog thereof, as discussed below) along with other components set out herein for treating a disease or disorder (i.e., indication) listed in Table 4, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a such a protein (or a homolog thereof, as discussed below) along with other components set out herein for treatment of a disease or disorder listed in Table 4.

In some embodiments, the compositions and methods of the invention provide for the delivery of one or more nucleic acids that bind to or otherwise act on an endogenous nucleic acid, such as endogenous mRNA or DNA, that alter the transcription, translation, secretion, or action of one or more proteins listed in Table 4 (or homologs thereof) or proteins useful in treating a disease or disorder (i.e., indication) listed in Table 4. For example, RNAi nucleic acids bind to endogenous mRNA, miRNA, or other RNAs, while guide RNA (gRNA) and CRISPR RNA (crRNA) respectively bind to DNA and act on DNA in the nucleus. Accordingly, in some embodiments, the compositions and methods of the invention provide for delivery of nucleic acids including one or more of RNAi nucleic acids, gRNA and crRNA to interfere with the transcription, translation, secretion, or action of one or more proteins listed in Table 4 (and homologs thereof) or proteins useful in treating a disease or disorder (i.e., indication) listed in Table 4, along with other components set out herein.

TABLE 4 Exemplary Indications and Related Proteins Indication Therapeutic Protein 3-Methylcrotonyl-CoA carboxylase deficiency Methylcrotonoyl-CoA carboxylase 3-Methylglutaconic aciduria Methylglutaconyl-CoA hydratase Actinic keratosis Acute intermittent porphyria Porphobilinogen deaminase Acute lymphocytic leukemia Acute myeloid leukemia Addison's disease Adenosine deaminase deficiency Adenosine deaminase Adrenoleukodystrophy ABCD1 Adrenomyeloneuropathy AIDS/HIV Alcohol use disorders Alkaptonuria Homogentisate 1,2-dioxygenase Allergic asthma Anti-IgE mAb Allergies (dermatitis, rhinitis) Alopecia areata Alpers' disease POLG Alpers-Huttenlocher syndrome Alpha 1-antitrypsin deficiency Alpha 1 protease inhibitor Alpha-mannosidosis Alpha-D-mannosidase Alport syndrome Alzheimer's disease Amyloid light-chain amyloidosis Amyotrophic lateral sclerosis (ALS) Anemia Erythropoietin Aortic valve stenosis Argininemia Arginase Argininosuccinic acidemia Argininosuccinate lyase Arrhythmogenic right ventricular dysplasia Autism Autosomal dominant and recessive progressive external ophthalmoplegia with mitochondrial DNA deletions Autosomal recessive polycystic kidney disease ARPKD Bacterial infections Basal cell carcinoma Batten disease Battenin + others B-cell chronic lymphocytic leukemia Becker muscular dystrophy Dystrophin Beta-thalassemia Beta globin Binge eating disorder Bipolar disorder Bladder cancer Blepharospasm, Cervical dystonia, Chronic migraine, Botulinum toxin more Bronchiolitis obliterans Brugada syndrome Buerger's disease CACNA1A CACNB4-related Episodic Ataxia Type 2 Cancer and depression Cancer and sexual dysfunction Cancer in pregnancy Carbamylphosphate synthetase deficiency Carbamylphosphate synthetase Carcinoma of the gallbladder Cardiomyopathy (diabetic) Cardiomyopathy (hypertrophic) Carnitine uptake defect SLC22A5 Catecholaminergic polymorphic ventricular tachycardia CDKL5-related Atypical Rett Syndrome Celiac disease Cellulitis Cerebrovascular disease Cervix uteri cancer Chronic fatigue syndrome Chronic graft versus host disease Chronic idiopathic urticaria Chronic immune thrombocytopenia Thrombopoietin Chronic kidney kisease Chronic liver disease Chronic lymphocytic leukemia Chronic myeloid leukemia Chronic pancreatitis Cirrhosis of the liver Citrullinemia, type I Argininosuccinate synthase Classic Rett Syndrome Classical galactosemia Galactose-1-phosphate uridylyltransferase Clostridium difficile associated diarrhea Clotting disorders COAD/COPD Cocaine addiction COL4A5-related disorders Cold contact urticaria Contraception, female Coronary artery diseases Corpus uteri cancer Corticobasal degeneration Crigler-Najjar syndrome UDP-glucuronosyltransferase Critical limb ischemia CTNS-related cystinosis Cutaneous lupus erythematosus Cutaneous neuroendocrine carcinoma (Merkel Cell) Cystic fibrosis CFTR Cystic fibrosis Deoxyribonuclease I Cystinosis Cystinosin Cystinuria SLC7A9 Dementia (Lewy body) Depression Diabetic foot infections Diabetic foot ulcer Diabetic peripheral neuropathy Diabetic ulcers Diarrhoeal diseases Diffuse large B-cell lymphoma DiGeorge syndrome Diverticulitis Drug use disorders Duchenne muscular dystrophy Dystrophin Dysarthria Dyskinesia (levodopa-induced) Early-onset autosomal dominant Alzheimer's disease Eczema Ehlers-Danlos syndrome, type 1 EIF2B1 EIF2B2 EIF2B3 EIF2B4 EIF2B5-related childhood ataxia with central nervous system hypomyelination/vanishing white matter Eosinophilic esophagitis Epilepsy Erectile dysfunction Erythropoietic protoporphyria Ferrochelatase Esophageal carcinoma Essential tremor Fabry disease Alpha galactosidase Familial adenomatous polyposis APC Familial chylomicronemia Lipoprotein lipase Familial dysbetalipoproteinemia Apolipoprotein E Familial isolated dilated cardiomyopathy Familial mediterranean fever Pyrin (MEFV) Familial melanoma Female infertility Follicle stimulating hormone Female sexual dysfunction Fibromyalgia FMR1-related disorders Fracture healing Fragile X Premature Ovarian Failure Syndrome Fragile X syndrome FMRP Fragile X-Associated Tremor/Ataxia Syndrome Friedreich's ataxia Frontotemporal dementia Fryns syndrome Galactocerebrosidase deficiencies GALE deficiency Galactose epimerase GALK deficiency Galactokinase GALT-related galactosemia Gastric cancer Gastroesophageal reflux disease Gaucher disease Glucocerebrosidase Gilbert syndrome UDP-glucuronosyltransferase Glioblastoma multiforme Glomerulonephritis Glutaric acidemia, type I Glutaryl-CoA dehydrogenase GM2 gangliosidosis HEXA, HEXB Gout Urate oxidase Graft versus host disease Growth hormone deficiency Growth hormone 1/Growth hormone 2 Head and neck cancer, Metastatic colorectal cancer Anti-EGFr mAb Hearing loss, adult onset Heart failure Hemachromatosis HFE protein Hemifacial spasm Hemolytic uremic syndrome Anti-complement factor C5 mAb Hemophilia A Factor VIII Hemophilia A, Hemophilia B Factor VII Hemophilia B Factor IX Hepatitis B, Hepatitis C Interferon alpha HER2+ breast cancer, gastric cancer Anti-HER2 mAb Hereditary angioedema C1 esterase inhibitor Hereditary hemorrhagic telangiectasia Hereditary hemorrhagic telangiectasia (AT) Hereditary spherocytosis Hidradenitis suppurativa Homocystinuria Cystathionine beta-synthase Homozygous familial hypercholesterolemia LDL receptor Hunter syndrome (MPS II) Iduronate-2-sulfatase Huntington disease Huntingtin Hurler syndrome (MPS I) Alpha-L iduronidase Hydrolethalus Hyperalgesia Hyperbilirubinemia Hyperhidrosis Hyperlipidemia Hypermethioninemia Methionine adenosyltransferase Hyperoxaluria, type I Serine-pyruvate aminotransferase Hypertension Hyperuricemia Hyponatremia Hypoparathyroidism Parathyroid hormone Hypophosphatasia TNSALP Idiopathic pulmonary fibrosis Iminoglycinuria Immunoglobulin deficiency Immunoglobulin Infection (adenovirus) Infection (anthrax prophylaxis) Infection (BK virus) Infection (Clostridium difficile prophylaxis) Infection (Dengue fever prophylaxis) Infection (Epstein-Barr virus) Infection (Hepatitis-D) Infection (Lyme disease prophylaxis) Infection (Smallpox virus) Infectious diseases vaccines Infectious antigen Inflammatory heart diseases Insomnia Interstitial cystitis Iron-deficiency anaemia Irritable bowel disease Ischaemic heart disease Isovaleric aciduria Isovaleric acid CoA dehydrogenase deficiency Jansky-Bielschowsky disease Juvenile Batten disease Juvenile Neuronal Ceroid Lipofuscinosis (JNCL) Juvenile rheumatoid arthritis TNF-alpha inhibitors Kennedy's disease (SBMA) Keratoconus Krabbe disease Galactocerebrosidase Leber's hereditary optic neuropathy NADH dehydrogenase Leiomyosarcoma Lennox-Gastaut syndrome Lesch-Nyhan syndrome Hypoxanthine phosphoribosyltransferase 1 Leukaemia Li-Fraumeni syndrome TP53 Lipoma Liposarcoma Liver cancer Long-chain 3-OH acyl-CoA dehydrogenase deficiency Long-chain-3-hydroxyacyl-CoA dehydrogenase Lower respiratory infections Lysosomal acid lipase deficiency Lysosomal acid lipase Macular degeneration Major depressive disorder Malignant fibrous histiocytoma Mantle cell lymphoma Maple syrup urine disease 3-methyl-2-oxobutanoate dehydrogenase Marfan syndrome FBN1 Maroteaux-Lamy syndrome (MPS VI) N-acetylgalactosamine 4-sulfatase Mastocytosis McArdle disease Muscle glycogen phosphorylase MECP2-related disorders MECP2-related Severe Neonatal Encephalopathy Medium-chain acyl-CoA dehydrogenase deficiency Acyl-CoA dehydrogenase Melanoma Anti-CTLA4 mAb Metachromatic leukodystrophy Arylsulfatase A Metastatic colorectal cancer, NSCLC, others Anti-VEGF mAb Methylmalonyl-CoA mutase deficiency Methylmalonyl-CoA mutase Migraine Mitochondrial oxidative phosphorylation disorders Morquio syndrome, type A (MPS IVA) Galactose 6-sulfate sulfatase Morquio syndrome, type B (MPS IVB) Beta-galactosidase Mouth and oropharynx cancers Multiple carboxylase deficiency Biotin-methylcrotonoyl-CoA-carboxylase ligase Multiple myeloma Multiple sclerosis Anti-VLA-4 mAb Multiple sclerosis Interferon beta Multiple system atrophy Myasthenia gravis Myelofibrosis Narcolepsy Neonatal bronchopulmonary dysplasia Neonatal infections Nephritis and nephrosis Neurofibromatosis, type 1 NF-1 Neuronal ceroid lipofuscinoses-related diseases Neutropenia G-CSF Niemann Pick disease, type A/B SMPD1 Niemann Pick disease, type C NPC1 Niemann-Pick disease Type C1 Nocturia Non-alcoholic fatty liver disease Non-Hodgkin lymphoma Anti-CD20 mAb Non-small cell lung cancer Notch-3 related cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) Obesity Ophthalmoparesis Opioid induced constipation Ornithine transcarbamylase deficiency Ornithine transcarbamylase Osteoarthritis Osteopetrosis Osteoporosis Anti-RANKL mAb Ovarian cancer Paget disease of bone Sequestosome 1 Pain Pancreatic carcinoma Panic disorder Parkinson disease Paroxysmal nocturnal hemoglobinuria Anti-complement factor C5 Mab Pediculosis capitis (head lice) Pelizaeus-Merzbacher disease Pemphigus vulgaris Peptic ulcer disease Peripheral neuropathy Peyronie's disease Phenylketonuria Phenylalanine hydroxylase Pneumococcal infection prophylaxis POLG-related sensory ataxic neuropathy Polycystic kidney disease Polycystic ovary syndrome Polycythaemia vera Polymerase G-related disorders Polymorphous light eruption Pompe disease Alpha glucosidase Porphyria cutanea tarda Uroporphyrinogen decarboxylase Post herpetic neuralgia Post-organ transplant Pouchitis PPM-X Syndrome Prader-Willi syndrome Preeclampsia Premature ejaculation Prematurity and low birth weight Primary ciliary dyskinesia Primary glomerular diseases Primary humoral immune deficiencies (e.g., CVID) Immunoglobulin Proctitis Progressive multifocal leukoencephalopathy Progressive supranuclear palsy Propionic acidemia Propionyl-CoA carboxylase Prostate cancer Psoriasis Anti-IL-12 & IL-23 mAb Psoriatic arthritis TNF-alpha inhibitors PTT-1 Pulmonary arterial hypertension Pulmonary arterial hypertension Raynaud's phenomenon Refractive errors Renal cell carcinoma Restless leg syndrome Retinitis pigmentosa Rheumatic heart disease Rheumatoid arthritis Anti-interleukin-6 (IL-6) mAb Rheumatoid arthritis T-cell costimulation blocker Rheumatoid arthritis TNF-alpha inhibitor Romano-Ward syndrome Rosacea Sanfilippo syndrome, type A (MPS IIIA) Heparan N-sulfatase Sanfilippo syndrome, type B (MPS IIIB) N-acetyl-alpha-D-glucosaminidase Santavuori-Haltia disease Schizophrenia Schnitzler syndrome Scleroderma SCN1A SCN1B-related seizure disorders Short-chain acyl-CoA dehydrogenase deficiency Butyryl-CoA dehydrogenase Sickle cell disease Hemoglobin SLC3A1-related disorders Small cell lung cancer SMN-1-related spinal muscular atrophy (SMA) Spinal muscular atrophy Survival motor neuron protein Squamous cell carcinoma of head and neck Stickler syndrome Stomach cancer Stroke prophylaxis Synovial sarcoma Systemic lupus erythematosus Anti-BAFF Systemic sclerosis Tetrahydrobiopterin-deficient hyperphenylalaninemia Tetrahydrobiopterin Thromboangiitis obliterans Thrombotic disorders Thyroid cancer TPP1 deficiencies Trachea, bronchus, lung cancers Tricuspid atresia TSC1 TSC2-related tuberous sclerosis Type 2 diabetes mellitus Glucagon-like peptide 1 (GLP-1) agonist Type 2 diabetes mellitus Insulin Tyrosinemia, type I Fumarylacetoacetase Ulcerative colitis Uterine fibroids Varicose veins Venous thromboembolism Very long-chain acyl-CoA dehydrogenase deficiency Long-chain-acyl-CoA dehydrogenase von Gierke's disease Glucose-6-phosphatase Von Hippel-Lindau disease pVHL Wegener granulomatosis Wilson disease Wilson disease protein X-Linked adrenal hypoplasia X-linked adrenoleukodystrophy X-linked agammaglobulinemia Bruton's tyrosine kinase

In some embodiments, the present invention is used to prevent, treat and/or cure a subject affected with a disease or disorder listed or associated with the proteins listed in Tables 1, 2, 3, or 4. In some embodiments, an mRNA encodes one or more of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), argininosuccinate synthetase (ASS1), Factor IX, survival motor neuron 1 (SMN1), or phenylalanine hydroxylase (PAH). In some embodiments, one or more of a RNAi nucleic acid, gRNA and crRNA bind to endogenous nucleic acid, such as RNA or DNA, to modulate one or more of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein, argininosuccinate synthetase (ASS1) protein, Factor IX, survival motor neuron 1 (SMN1) protein, phenylalanine hydroxylase (PAH) or a protein or gene that modulates such proteins.

EXAMPLES Example 1. Synthesis of Thioacetate Starting Material (Scheme 8)

In a typical procedure, 25 g of potassium thioacetate is dissolved in 200 mL of ACS grade acetone and cooled in an ice bath. To this, 27.4 g of 2-bromoethanol is added to the cooled solution (the container is then rinsed with ^(˜)50 mL of acetone to ensure maximum transfer). The reaction is stirred for 24 hours, allowing the bath to come to room temperature. The solution is then filtered by gravity before removing the solvent by rotovap to give crude yellow oil. The resulting oil is dissolved in 500 mL of DCM, washed twice with 200 mL of water and dried over sodium sulfate before decanting and evaporating to give crude product (75-85% yield). The product is then purified by column chromatography (EtOAc:Heptane). ¹H NMR: 2.37 ppm (s, 3H); 2.64 ppm (broad —OH, 1H); 3.08 ppm (t, 2H); 3.75 ppm (t, 2H).

Example 2. Production of Cyclophospholane Precursor

Phosphoryl trichloride and TEA were dissolved in 30 mL of DCM and cooled to −35° C. (IPA/CO₂ bath). The diol was added slowly keeping the temp below −20° C. and the product was brought to room temperature and mixed for at least 1 hour. The solution was cooled again in IPA/CO₂. A TEA/thioester mixture in 3 volumes of DCM was added to the reaction slowly, keeping the temperature below −20° C. The solution was then allowed to come to room temperature while mixing overnight The solution was filtered the filtrate was then concentrated by rotovap. The resulting oil was dissolved in DCM and rinsed with bicarbonate/water. The organic layer was dried by MgSO₄, filtered and concentrated to give quantitative crude yield. ¹H NMR consistent with expected structure.

Example 3. Production of Poly(Phosphoester) Using Ring-Opening Polymerization (ROP)

In a 50 mL round bottom flask, 2 g of monomer were charged into 20 mL of anhydrous toluene and 7.6 mg of benzyl alcohol. The mixture was stirred for 15 minutes under nitrogen and 13.4 mg of DBU was added via syringe and allowed to stir for 24 hours. After this time, the reaction was quenched with excess acetic acid. The resulting polymer was precipitated with 20 volumes of diethyl ether to give 20% yield of polymer with Mw of 900 Da.

Example 4. Production of Poly(Phosphoester) Using Ring-Opening Polymerization (ROP) as a One-Pot Reaction Using Pendant Amine

A 100 mL round bottom flask was charged with 25 mL of DCM and sealed under positive nitrogen pressure before adding 2 g of POCl₃. The reaction mixture was cooled in IPA/CO₂ bath for 30 minutes before adding premixed thioester/DCM/TEA solution (1.6 g/5 mL/1.4 g, respectively) slowly, never letting the solution rise above −20° C. After addition, stirring was continued and the reaction mixture was brought to room temperature for at least 1 hour. The solution was cooled in IPA/CO₂ bath for at least 30 minutes and a premix of diol/DCM/TEA solution (1.57 g/3 mL/2.6 g respectively) was added slowly. The reaction stirred for up to 72 hours at room temperature. Upon completion, the reaction was filtered and the filtered solution was then added to 10 volumes of MTBE slowly with constant stirring. The solids were removed by filtration and the filtrate was concentrated under reduced pressure. The concentrated solution was precipitated in 20 volumes of cold THF and decanted. The product was re-dissolved in MeOH (5 volumes) and precipitated into THF again. The product was filtered, and dried under vacuum to constant mass (0.5 g, 10% yield).

Example 5. Production of Poly(Phosphoester) Using Ring-Opening Polymerization (ROP) as a One-Pot Reaction Using Amine within Polymer Backbone

A 100 mL round bottom flask was charged with 25 mL of DCM and sealed under positive nitrogen pressure before adding 2 g POCl₃. The reaction mixture was cooled in IPA/CO₂ bath for 30 minutes before adding the premixed thioester/DCM/TEA solution (1.6 g/5 mL/1.4 g, respectively) slowly, never letting the solution rise above −20° C. After addition, stirring was continued and the reaction mixture was brought to room temperature for at least 1 hour. After this time, the solution was cooled in IPA/CO₂ bath for at least 30 minutes and premixed diol/DCM/TEA solution (1.57 g/3 mL/2.6 g respectively) was added slowly. The reaction stirred for up to 72 hours at room temperature and was then filtered. The filtered solution was then added to 10 volumes of MTBE slowly with constant stirring and the resulting solids were removed by filtration. The filtrate was concentrated under reduced pressure. The resulting concentrated solution precipitated in 20 volumes of cold THF. The THF was decanted and the product was dissolved in MeOH (5 volumes) and precipitated into THF again. After decantation, the solid was dried under vacuum to constant mass (1.2 g 22% yield).

Example 6. Synthesis of Dinitrophenol Thioacetate Phosphate Precursor Starting Material (One-Pot Example)

A 100 mL round bottom flask was charged with 75 mL of DCM and 3 g of POCl₃ and sealed under positive nitrogen pressure. The reaction mixture was cooled in an ice bath for 30 minutes before slowly adding a premixed thioester/DCM/TEA solution (2.44 g/10 mL/1.8 g, respectively) slowly, never letting the solution rise above 20° C. After addition, stirring was continued while the reaction mixture was brought to room temperature for at least 1 hour. After this time, the solution was cooled in ice bath for at least 30 minutes. Upon cooling, a premixed phenol/DCM/TEA solution (5.7 g/10 mL/4 g respectively) was slowly over the course of an hour and the reaction was stirred for up to 72 hours at ambient temperature. Upon completion, the reaction was filtered and the filtrate was concentrated under reduced pressure. The resulting crude material was purified by silica gel chromatography with DCM/MeOH mixture (56% yield).

Example 7. Synthesis of Dinitrophenol Chlorophosphate Starting Material

In a glove bag, 16 g bisphenol phosphoryl chloride was charged to 100 mL round bottom flask. 25 mL of carbon tetrachloride was add to the flask and the solution mixed to dissolve the solid. The sealed vessel was transferred to a cooled in an ice bath for 30 minutes under N₂ and stirring was performed. 20 mL of a 3:7 HNO₃:H₂SO₄ solution was added slowly through a drop funnel over 1 hour and allowed to stir overnight (16 hours). After this time, the reaction was washed with 200 mL of DCM, and the organic layer was separated and quenched with anhydrous sodium carbonate under N₂. The solution was then filtered and the solvent evaporated to a constant mass. The product was used as-is in subsequent reactions (21 g crude, quantitative). ¹H NMR consistent with expected structure.

Example 8. Alternate Synthesis of Dinitrophenol Thioacetate Precursor

A 250 mL round bottom flask was charged with 10 g of dinitrophenol chlorophosphate with 100 mL of DCM under N₂. The reaction mixture was cooled in an IPA/CO₂ bath and mixed until a homogenous solution was achieved. Slow addition of a premixed thioester/DCM/TEA solution (2.1 g/10 mL/2.2 g) to the reaction vessel performed and mixed slowly over and hour. The reaction was brought to room temperature while stirring under N₂ overnight. After this time, the reaction solution was evaporated and the residue was dissolved in 100 mL of carbon tetrachloride, filtered, and evaporated again to a constant mass (6.5 g, 80% yield). NMR was consistent with desired product.

Example 9. Alternate Production of Poly(Phosphoester)

A 100 mL round bottom flask was charged with 25 mL of DCM and sealed under positive nitrogen pressure before adding 2 g POCl₃. The reaction mixture was cooled in IPA/CO₂ bath for 30 minutes before adding premixed thioester/DCM/TEA solution (1.6 g/5 mL/1.4 g, respectively) slowly, never letting the solution rise above −20° C. After addition of the mixture, stirring was continued and the reaction mixture was brought to room temperature for at least 1 hour. The solution as cooled solution in IPA/CO₂ bath for at least 30 minutes. Upon cooling, a premixed diol/DCM/TEA solution (1.57 g/3 mL/2.6 g respectively) was added and the resulting mixture was stirred for up to 72 hours at room temperature. After a given time, the reaction was filtered and the filtrate was added to 10 volumes of MTBE slowly with constant stirring. The resulting solids were removed by filtration and the filtrate was concentrated under reduced pressure. The concentrated solution was precipitated in 20 volumes of cold THF and the solvent was decanted. The product was re-dissolved in MeOH (5 volumes) and precipitated into THF again. The THF was decanted and the product was dried under vacuum to constant mass (0.4 g, 32% yield).

Example 10. Alternate Production of Poly(Phosphoester)

TABLE A Reaction # Monomer Amine Conditions MW Notes 074

TEA DCM 12.8 kDa Dialyzed 079A

TEA DCM   24 kDa Dialyzed 079B

Polymer- bound scavenger (resin) DCM   24 kDa Dialyzed 086

Polymer- bound scavenger (resin) DCM   21 kDa Dilute in MeOH (filter), Remove and recrystallize in THF 087A

TEA DCM   45 kDa MTBE precipitation (filter), MeOH/THF recrystallization 087B

TEA DCM   32 kDa Acidify reaction mixture, Toluene addition (oil out) MeOH/THF recrystallization 088

TEA DCM   69 kDa MeOH/Brine THF Precipitation 089

TEA DCM   24 kDa MeOH/Brine THF extracted 090

TEA DCM   7 kDa MeOH/EtOAc recrystallization

Example 11. Synthesis of Branched Poly(Phosphoester)

POCl₃ was dissolved in dichloromethane (5 mL) in a 100 mL round bottom flask. The solution was cooled with IPA/CO2 before adding the diol/TEA/DCM solution slowly. The reaction solution was stirred for 72 hours, quenched with MeOH/ACN and the solvent was removed under reduced pressure to dryness yielding a crude yellow material. A portion of the material was dissolved and dialyzed against a 50/50 MeOH/phosphate buffer solution for 24 hours resulting in a polymer with an approximate molecular weight of 27 kDa.

Synthesis of Alternative Imidazole-Based Diol Monomers.

Exemplary imidazole-based diols are provided in Scheme 19.

Scheme 20 describes an exemplary synthesis of diol monomer 301.

Scheme 21 describes an exemplary synthesis of diol monomer 302.

Scheme 22 describes an exemplary synthesis of diol monomer 303.

Scheme 23 describes an exemplary synthesis of diol monomer 304.

Example 12. Synthesis of Diol Monomer 305

Scheme 24 describes an exemplary synthesis of diol monomer 305.

N-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)propanamide (3)

To the mixture of 3-(1-trityl-1H-imidazol-4-yl)propanoic acid (1) (10.2 g, 0.027 mol, 1.0 equiv) and HATU (10.2 g, 0.027 mol, 1.0 equiv) in DCM (200 mL) was added DIPEA (14 mL, 3.0 equiv), the resulting mixture was stirred 5 min. at room temperature, and then (2,2-dimethyl-1,3-dioxolan-4-yl)methanamine (2) (3.5 g, 0.027 mol, 1.0 equiv) was added. The mixture was stirred overnight at room temperature. The reaction mixture was washed with Brine (100 mL×3), and dried with anhydrous sodium sulfate. After the filtration, the organic solvent was evaporated under vacuum and the residue was purified with an ISCO (330 g column), eluted with 0-100% MeOH in DCM, collected 13 g (98%) of the desired product 3.

(2,2-dimethyl-1,3-dioxolan-4-yl)methyl methanesulfonate (5)

To the solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methanol (4) (25 g, 0.18 mol) in DCM (100 mL) was added TEA (30 mL, 0.32 mol) followed by the addition of MSCl (16.3 mL, 0.15 mol) with ice bath cooling. The resulting mixture was stirred for 2 h. at room temperature. The reaction mixture was then washed with cold water (100 mL×3), dried with anhydrous magnesium sulfate. After filtration, the organic solvent was evaporated under vacuum and collected 37.2 g of crude product (5) that was used for the next step without further purification.

4-(azidomethyl)-2,2-dimethyl-1,3-dioxolane (6)

To the solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methyl methanesulfonate (7) (37.2 g, 0.18 mol) in DMF (100 mL) was added a solution of NaN₃ (16 g, 0.24 mol, 1.3 equiv) in water (100 mL). The resulting mixture was heated overnight at 80-90° C. After the reaction was cooled to room temperature, brine (400 mL) was added. Ether was used to extract (200 mL×4), the combined ether was washed with brine (200 mL×2) and water (200 mL×2), dried with anhydrous sodium sulfate. After the filtration, the organic solvent was evaporated under vacuum and collected 21.7 g of crude product (6) that was used for the next step without further purification

(2,2-dimethyl-1,3-dioxolan-4-yl)methanamine (2)

To the solution 4-(azidomethyl)-2,2-dimethyl-1,3-dioxolane (6) (20 g) in MeOH (200 mL), Pd/C (10%, 5 g) was added, the resulting mixture was purged with H₂ three times and stirred under H₂ balloon overnight. The reaction mixture was passed through a Celite pad, washed with methanol. The filtrate was concentrated and purified with an ISCO (330 g column, eluted with 0-10% MeOH in DCM, collected 5.8 g (17%) desired product 2.

N-(2,3-dihydroxypropyl)-3-(1H-imidazol-4-yl)propanamide (Target 305)

To the solution of N-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)propanamide (3) (13 g) in DCM (25 mL) and MeOH (25 mL) was added AcOH (100 mL). The resulting reaction mixture was heated overnight at 70-80° C. The reaction solution was cooled to room temperature and concentrated under vacuum. MeOH (50 mL) was added into the residue, the pH was adjusted to 9-10 with 2M NaOH aqueous solution. After the filtration, the filtrate was purified with an ISCO, 330 g C-18 column, eluted with 0-100% MeOH/H₂O, collected 2.5 g (44%) of pure desired product (Target 305.)

Example 13. Synthesis of Diol Monomer 306

Scheme 25 describes an exemplary synthesis of diol monomer 306.

N-((2,2-dimethyl-1,3-dioxan-5-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)propanamide (3)

To the mixture of 3-(1-trityl-1H-imidazol-4-yl)propanoic acid (1) (15 g, 0.039 mol, 1.0 equiv) and HATU (15 g, 0.039 mol, 1.0 equiv) in DCM (400 mL), DIPEA (20 mL, 3.0 equiv) was added, the resulting mixture was stirred 5 min. at room temperature, and (2,2-dimethyl-1,3-dioxan-5-yl)methanamine (4) (5.7 g, 0.039 mol, 1.0 equiv) was added. The resulting mixture was stirred overnight at room temperature. The reaction mixture was washed with Brine (100 mL×3), and dried with anhydrous sodium sulfate. After the filtration, the organic solvent was evaporated under vacuum and the residue was purified with ISCO (330 column), eluted with 0-100% MeOH in DCM, collected 17.6 g (88%) desired product 3.

N-(3-hydroxy-2-(hydroxymethyl)propyl)-3-(1H-imidazol-4-yl)propanamide (Target 306)

To the solution of N-((2,2-dimethyl-1,3-dioxan-5-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)propanamide (3) (17.6 g) in DCM (50 mL) and MeOH (50 mL) was added AcOH (100 mL). The resulting reaction mixture was heated overnight at 70-80° C. The reaction solution was cooled to room temperature and concentrated under vacuum. MeOH (50 mL) was added into the residue, the pH was adjusted to 9-10 with 2M NaOH aqueous solution. After the filtration, the filtrate was purified with ISCO, 330 g C-18 column, eluted with 0-100% MeOH/H₂O, collected 4.4 g (56%) pure desired product (Target 306.)

Example 14. Synthesis of Diol Monomer 307

Scheme 26 describes an exemplary synthesis of diol monomer 307.

(E)-N-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)acrylamide (3)

To the mixture of (E)-3-(1-trityl-1H-imidazol-4-yl)acrylic acid (1) (14.5 g, 0.038 mol, 1.0 equiv) and HATU (14.5 g, 0.038 mol, 1.0 equiv) in DCM (400 mL), DIPEA (19.6 mL, 3.0 equiv) was added, the resulting mixture was stirred 5 min. at room temperature, and (2,2-dimethyl-1,3-dioxan-5-yl)methanamine (4) (5.0 g, 0.038 mol, 1.0 equiv) was added. The resulting mixture was stirred overnight at room temperature. The reaction mixture was washed with Brine (100 mL×3), and dried with anhydrous sodium sulfate. After filtration the organic solvent was evaporated under vacuum and the residue was purified with ISCO (330 g column), eluted with 0-100% MeOH in DCM, collected 17.1 g (90%) desired product 5.

(E)-N-(2,3-dihydroxypropyl)-3-(1H-imidazol-4-yl)acrylamide (Target 307)

To the solution of (E)-N-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)acrylamide (3) (17.1 g) in DCM (50 mL) and MeOH (50 mL) was added AcOH (100 mL). The resulting reaction mixture was heated overnight at 70-80° C. The reaction solution was cooled to room temperature and concentrated under vacuum. MeOH (50 mL) was added into the residue, the pH was adjusted to 9-10 with 2M NaOH aqueous solution. After filtration the filtrate was purified with an ISCO, 330 g C-18 column, eluted with 0-100% MeOH/H₂O, collected 2.0 g (27%) pure desired product (Target 307.)

Example 15. Synthesis of Diol Monomer 308

Scheme 27 describes an exemplary synthesis of diol monomer 308.

Synthesis of (E)-3-(1-trityl-1H-imidazol-4-yl)acrylic Acid (3)

To the mixture of urocanic acid (1) (21.3 g, 0.154 mol, 1.0 equiv) and trityl chloride (2) (43 g, 0.154 mol, 1.0 equiv) in anhydrous DMF (350 mL) was added TEA (64.8 mL, 0.462 mol, 3.0 equiv). The mixture was stirred overnight at room temperature. CHCl₃ (500 mL) was added and the solution washed with Brine. The aqueous was further extracted with DCM and the combined organic phases were combined and dried over anhydrous sodium sulfate. After filtration the organic solvent was evaporated under vacuum and the residue was purified with an ISCO (330 column), eluted with 0-15% MeOH in DCM, collected 28.4 g (48%) of the desired product 3.

(E)-N-((2,2-dimethyl-1,3-dioxan-5-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)acrylamide (5)

To the mixture of (E)-3-(1-trityl-1H-imidazol-4-yl)acrylic acid (3) (15 g, 0.039 mol, 1.0 equiv) and HATU (15 g, 0.039 mol, 1.0 equiv) in DCM (400 mL), was added DIPEA (20 mL, 3.0 equiv) and the resulting mixture was stirred 5 min. at room temperature and then (2,2-dimethyl-1,3-dioxan-5-yl)methanamine (4) (5.7 g, 0.039 mol, 1.0 equiv) was added. The mixture was stirred overnight at room temperature. The reaction mixture was then washed with Brine (100 mL×3), and dried with anhydrous sodium sulfate. After the filtration, the organic solvent was evaporated under vacuum and the residue was purified with ISCO (330 column), eluted with 0-100% MeOH in DCM, collected 18.2 g (91%) of the desired product 5.

(2,2-dimethyl-1,3-dioxan-5-yl)methyl methanesulfonate (7)

To the solution of (2,2-dimethyl-1,3-dioxan-5-yl)methanol (6) (20 g, 0.137 mol, 1.0 equiv) in DCM (400 mL), was added TEA (45.4 mL, 0.32 mol, 2.3 equiv), followed by the addition of MSCl (11.7 mL, 0.15 mol) with ice bath cooling. The resulting mixture was stirred for 2 h. at room temperature. The reaction mixture was washed with cold water (100 mL×3), dried with anhydrous magnesium sulfate. After the filtration, the organic solvent was evaporated under vacuum and collected 27 g of crude product (7) that was used for the next step without further purification.

5-(azidomethyl)-2,2-dimethyl-1,3-dioxane (8)

To the solution of (2,2-dimethyl-1,3-dioxan-5-yl)methyl methanesulfonate (7) (27 g, 0.12 mol) in DMF (100 mL) was added a solution of NaN₃ (11.7 g, 0.18 mol, 1.5 equiv) in water (100 mL). The resulting mixture was heated overnight at 80-90° C. After the reaction was cooled to room temperature, brine (400 mL) was added. Ether was used to extract organic material (200 mL×4) and the combined ether extracts were washed with brine (200 mL×2) and water (200 mL×2) and dried with anhydrous sodium sulfate. After filtration, the organic solvent was evaporated under vacuum and collected 15.7 g of crude product (8) that was used for the next step without further purification.

(2,2-dimethyl-1,3-dioxan-5-yl)methanamine (4)

To the solution of 5-(azidomethyl)-2,2-dimethyl-1,3-dioxane (8) (15.7 g) in MeOH (200 mL) was added Pd/C (10%, 4.5 g) and the resulting mixture was purged with H₂ gas three times and stirred under a H₂ balloon overnight. The reaction mixture was passed through a Celite pad, and the pad was washed with methanol. The filtrate was concentrated and collected 11.5 g desired product 4 that was used for the next step without further purification.

(E)-N-(3-hydroxy-2-(hydroxymethyl)propyl)-3-(1H-imidazol-4-yl)acrylamide (Target 308)

To the solution of (E)-N-((2,2-dimethyl-1,3-dioxan-5-yl)methyl)-3-(1-trityl-1H-imidazol-4-yl)acrylamide (5) (18.2 g) in DCM (50 mL) and MeOH (50 mL) was added AcOH (100 mL). The resulting reaction mixture was heated overnight at 70-80° C. The reaction solution was cooled to room temperature and concentrated under vacuum. MeOH (50 mL) was added into the residue and the pH was adjusted to 9-10 with 2M NaOH aqueous solution. After filtration, the filtrate was purified with by ISCO, 330 g C-18 column, eluted with 0-100% MeOH/H₂O, collected 3.9 g (48%) pure desired product (Target 308.)

Examples of mRNA-Poly(Phosphoester) Nanoparticles Example 16. Production of Human Argininosuccinate Synthetase (hASS1) Poly(Phosphoester) Nanoparticles

A portion of poly(phosphoseter) 086 was dissolved in water and the pH was adjusted to 6.5. The solution was treated with a 1 mL solution of hASS1 mRNA (1 mg) and mixed resulting in a nanoparticle formation (N/P=10) with an average particle size of 89 nm.

Example 17. Production of hASS1 Poly(Phosphoester) Nanoparticles

A portion of poly(phosphoseter) 087A was dissolved in water and the pH was adjusted to 6.5. The solution was treated with a 1 mL solution of hASS1 mRNA (1 mg) and mixed resulting in a nanoparticle formation (N/P=10) with an average particle size of 92 nm. Example 21. Production of hASS1 poly(phosphoester) nanoparticles

A portion of poly(phosphoseter) 087B was dissolved in water and the pH was adjusted to 6.5. The solution was treated with a 1 mL solution of hASS1 mRNA (1 mg) and mixed resulting in a nanoparticle formation (N/P=10) with an average particle size of 90 nm.

Codon-Optimized Sequence for Human Argininosuccinate Synthetase (ASS1) mRNA:

[SEQ ID NO.: 1] XAUGAGCAGCAAGGGCAGCGUGGUGCUGGCCUACAGCGGCGGCCUGGACAC CAGCUGCAUCCUGGUGUGGCUGAAGGAGCAGGGCUACGACGUGAUCGCCUA CCUGGCCAACAUCGGCCAGAAGGAGGACUUCGAGGAGGCCCGCAAGAAGGC CCUGAAGCUGGGCGCCAAGAAGGUGUUCAUCGAGGACGUGAGCCGCGAGUU CGUGGAGGAGUUCAUCUGGCCCGCCAUCCAGAGCAGCGCCCUGUACGAGGA CCGCUACCUGCUGGGCACCAGCCUGGCCCGCCCCUGCAUCGCCCGCAAGCA GGUGGAGAUCGCCCAGCGCGAGGGCGCCAAGUACGUGAGCCACGGCGCCAC CGGCAAGGGCAACGACCAGGUGCGCUUCGAGCUGAGCUGCUACAGCCUGGC CCCCCAGAUCAAGGUGAUCGCCCCCUGGCGCAUGCCCGAGUUCUACAACCG CUUCAAGGGCCGCAACGACCUGAUGGAGUACGCCAAGCAGCACGGCAUCCC CAUCCCCGUGACCCCCAAGAACCCCUGGAGCAUGGACGAGAACCUGAUGCA CAUCAGCUACGAGGCCGGCAUCCUGGAGAACCCCAAGAACCAGGCCCCCCC CGGCCUGUACACCAAGACCCAGGACCCCGCCAAGGCCCCCAACACCCCCGA CAUCCUGGAGAUCGAGUUCAAGAAGGGCGUGCCCGUGAAGGUGACCAACGU GAAGGACGGCACCACCCACCAGACCAGCCUGGAGCUGUUCAUGUACCUGAA CGAGGUGGCCGGCAAGCACGGCGUGGGCCGCAUCGACAUCGUGGAGAACCG CUUCAUCGGCAUGAAGAGCCGCGGCAUCUACGAGACCCCCGCCGGCACCAU CCUGUACCACGCCCACCUGGACAUCGAGGCCUUCACCAUGGACCGCGAGGU GCGCAAGAUCAAGCAGGGCCUGGGCCUGAAGUUCGCCGAGCUGGUGUACAC CGGCUUCUGGCACAGCCCCGAGUGCGAGUUCGUGCGCCACUGCAUCGCCAA GAGCCAGGAGCGCGUGGAGGGCAAGGUGCAGGUGAGCGUGCUGAAGGGCCA GGUGUACAUCCUGGGCCGCGAGAGCCCCCUGAGCCUGUACAACGAGGAGCU GGUGAGCAUGAACGUGCAGGGCGACUACGAGCCCACCGACGCCACCGGCUU CAUCAACAUCAACAGCCUGCGCCUGAAGGAGUACCACCGCCUGCAGAGCAA GGUGACCGCCAAGUGAY 5′ and 3′ UTR Sequences X (5′ UTR Sequence) = [SEQ ID NO.: 2] GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGA CACCGGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGG AUUCCCCGUGCCAAGAGUGACUCACCGUCCUUGACACG Y (3′ UTR Sequence) = [SEQ ID NO.: 3] CGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGU UGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAA GCU OR (SEQ ID NO.: 4) GGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUU GCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAA GCU

While a number of embodiments of this invention have been described, it is apparent that the basic examples may be altered to provide other embodiments that utilize the compounds, methods, and processes of this invention. Therefore, it will be appreciated that the scope of this invention is to be defined by the appended claims rather than by the specific embodiments that have been represented by way of example herein. 

We claim:
 1. A polymer comprising a repeating unit that is

or a pharmaceutically acceptable salt thereof, wherein: each instance of R¹ independently is —H or a C₁-C₂₀ aliphatic group; each instance of R² independently is —H or a C₁-C₂₀ aliphatic group; each instance of R³ independently is —H or a C₁-C₂₀ aliphatic group; X is independently S or O; n1 is an integer from 1 to 6; n2 is an integer from 1 to 6; R is N or CH; and L is C₁₋₆ alkyl or a protonatable group provided that when R is CH then L is a protonatable group, wherein the protonatable group comprises at least one 1° amino, 2° amino, 3° amino or imidazolyl group.
 2. The polymer of claim 1, wherein R¹ independently is —H, —CH₃, or —CH₂CH₃.
 3. The polymer of claim 1, wherein R² independently is —H, —CH₃, or —CH₂CH₃.
 4. The polymer of claim 1, wherein R³ independently is —H, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, CH(CH₃)₂, or C(CH₃)₃.
 5. The polymer of claim 1, wherein X is independently 0 or S.
 6. The polymer of claim 1, wherein the polymer is a homopolymer.
 7. The polymer of claim 1, wherein the polymer is a copolymer.
 8. The polymer of claim 7, wherein the polymer is an alternating copolymer.
 9. The polymer of claim 7, wherein the polymer is a block copolymer.
 10. The polymer of claim 1, wherein: R is N; and L is C₁₋₆ alkyl; R is CH; and L is a protonatable group; or R is N; and L is a protonatable group.
 11. The polymer of claim 1, wherein: L is a protonatable group such that at least one conjugate acid of L-H has a pKa in water of about 4.5 to about 11.5.
 12. The polymer of claim 1, comprising a moiety T that is a targeting group.
 13. A pharmaceutical composition comprising: the polymer of claim 1; and one or more polynucleotides.
 14. The pharmaceutical composition of claim 13, wherein the one or more polynucleotides comprise RNA, wherein the RNA is selected from the group consisting of mRNA, siRNA, snoRNA, microRNA, gRNA, crRNA, and combinations thereof.
 15. The pharmaceutical composition of claim 14, wherein the RNA is mRNA.
 16. The pharmaceutical composition of claim 15, wherein the mRNA encodes a peptide, a polypeptide, or an enzyme.
 17. The pharmaceutical composition of claim 15, wherein the mRNA encodes cystic fibrosis transmembrane conductance regulator (CFTR) protein for use in the delivery to or treatment of the lung of a subject or a lung cell.
 18. The pharmaceutical composition of claim 15, wherein the mRNA encodes ornithine transcarbamylase (OTC) protein for use in the delivery to or treatment of the liver of a subject or a liver cell.
 19. The pharmaceutical composition of claim 15, wherein the mRNA encodes a peptide or polypeptide for use in vaccine, and wherein the mRNA encodes an antigen.
 20. A method of treating a disease in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of claim
 13. 21. A polymer comprising a repeating unit that is

or a pharmaceutically acceptable salt thereof, wherein: each instance of R¹ independently is —H or a C₁-C₂₀ aliphatic group; each instance of R² independently is —H or a C₁-C₂₀ aliphatic group; each instance of R³ independently is —H or a C₁-C₂₀ aliphatic group; X is independently S or O; n2 is an integer from 1 to 6; R is N or CH; and L is C₁₋₆ alkyl or a protonatable group provided that when R is CH then L is a protonatable group, wherein the protonatable group comprises at least one 1° amino, 2° amino, 3° amino or imidazolyl group.
 22. A homopolymer comprising a repeating group selected from the group consisting of: 